Combination comprising a tim-3 inhibitor and a hypomethylating agent for use in treating myelodysplastic syndrome or chronic myelomonocytic leukemia

ABSTRACT

Combination therapies comprising TIM-3 inhibitors are disclosed. The combinations can be used to treat cancerous conditions and disorders, including hematologic cancers.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/962,653, filed on Jan. 17, 2020, U.S. Provisional Application No.63/061,001, filed on Aug. 4, 2020, and U.S. Provisional Application No.63/125,691, filed on Dec. 15, 2020. The contents of the aforementionedapplications are hereby incorporated by reference in their entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jan. 11, 2021, isnamed C2160-7026WO_SL.txt and is 59,558 bytes in size.

BACKGROUND

Myelodysplastic syndromes (MDS) correspond to a heterogeneous group ofhematological malignancies associated with impaired bone marrowfunction, ineffective hematopoiesis, elevated bone marrow blasts, andpersistent peripheral blood cytopenias. Anemia is one of the most commonsymptoms of MDS and as a result, most patients with MDS undergo at leastone red blood cell transfusion. MDS can also progress to acute myeloidleukemia (AML) (Heaney and Golde (1999) N. Engl, J. Med.340(21):1649-60). Although progression to AML can lead to death inpatients with MDS, MDS-related deaths can also result from cytopeniasand marrow failure in the absence of leukemic transformation. Prognosisof MDS is typically determined using the revised InternationalPrognostic Scoring System (IPSS-R), which considers the percentage ofbone marrow blasts, the number of cytopenias, and bone marrowcytogenetics. Patients with untreated MDS are classified into fiveIPSS-R prognostic risk categories: very low, low, intermediate, high andvery high, (Greenberg et al. (2012) Blood 108(11):2623).

Chronic myelomonocytic leukemia (CMML) is a clonal hematopoietic stemcell disorder with overlapping features of myelodysplastic syndromes andmyeloproliferative neoplasms, with an inherent risk for leukemictransformation (Patnaik et al. (2018) Am J Hematol 93(6)824-840). CMMLis characterized by the presence of sustained (>3 month) peripheralblood monocytosis along with dysplastic features in the bone marrow. Apatient with CMML is classified into three different subgroups based onpercentage of peripheral blasts and marrow blasts present. CMML-0corresponds, e.g., to about ≤2% peripheral blasts and about <5% marrowblasts, CMML-1 corresponds, e.g., to 2-4% peripheral blasts and about5-9% marrow blasts, and CMML-2 corresponds, e.g., to >5% peripheralblasts and 10-19% marrow blasts.

Prognosis is poor and life expectancy is short in intermediate, high, orvery high risk MDS, and chronic myelomonocytic leukemia 2 (CMML-2)patients. The current standard of care is the use of a hypomethylatingagent, chemotherapy, and/or hematopoietic stem cell transplant (HSCT).HSCT is the only curative option. However, only a minority of MDS orCMML patients are candidates for HSCT and intensive chemotherapy(Steensma (2018) Blood Cancer J 8(5): 47; Platzbecker (2019) Blood133(10): 1096-1107; Itzykson et al. (2018) HemaSphere 2(6):150).Complete remission is only reported in a minority of patients treated byazacitidine alone, and clinical benefits of this drug are frequentlytransient. When treatment fails, additional treatment options arelimited. Despite the fact that single-agent hypomethylating agents areavailable for the treatment of patients with higher risk MDS and CMML-2,alternative treatment strategies are needed.

SUMMARY

Disclosed herein, at least in part, are combinations comprisinginhibitors of T-cell immunoglobulin domain and mucin domain 3 (TIM-3).In some embodiments, the combination comprises an antibody molecule(e.g., a humanized antibody molecule) that binds to TIM-3 with highaffinity and specificity. In some embodiments, the combination furthercomprises a hypomethylating agent. Pharmaceutical compositions and doseformulations relating to the combinations described herein are alsoprovided. The combinations described herein can be used to treat orprevent disorders, such as cancerous disorders (e.g., hematologicalcancers). Thus, methods, including dosage regimens, for treating variousdisorders using the combinations are disclosed herein.

Accordingly, in one aspect, the disclosure features a method of treatinga hematological cancer, e.g., a myelodysplastic syndrome (MDS) in asubject, comprising administering to the subject a combination of aTIM-3 inhibitor and a hypomethylating agent.

In some embodiments, the TIM-3 inhibitor comprises an anti-TIM-3antibody molecule. In some embodiments, the TIM-3 inhibitor comprises ananti-TIM-3 antibody molecule. In some embodiments, the TIM-3 inhibitorcomprises MBG453, TSR-022, LY3321367, Sym023, BGB-A425, INCAGN-2390,MBS-986258, RO-7121661, BC-3402, SHR-1702, or LY-3415244. In someembodiments, the TIM-3 inhibitor comprises MBG453. In some embodiments,the TIM-3 inhibitor is administered at a dose of about 700 mg to about900 mg. In some embodiments, the TIM-3 inhibitor is administered at adose of about 800 mg. In some embodiments, the TIM-3 inhibitor isadministered at a dose of about 300 mg to about 500 mg. In someembodiments, the TIM-3 inhibitor is administered at a dose of about 400mg. In some embodiments, the TIM-3 inhibitor is administered once everyfour weeks. In some embodiments, the TIM-3 inhibitor is administered onday 8 of a 28-day cycle. In some embodiments, the TIM-3 inhibitor isadministered once every two weeks. In some embodiments, the TIM-3inhibitor is administered on day 8 and day 22 of a 28-day cycle. In someembodiments, the TIM-3 inhibitor is administered once every four weeks.In some embodiments, the TIM-3 inhibitor is administered intravenously.In some embodiments, the TIM-3 inhibitor is administered intravenouslyover a period of about 15 minutes to about 45 minutes. In someembodiments, the TIM-3 inhibitor is administered intravenously over aperiod of about 30 minutes.

In some embodiments, the hypomethylating agent comprises azacitidine,decitabine, CC-486 or ASTX727. In some embodiments, the hypomethylatingagent comprises azacitidine. In some embodiments, the hypomethylatingagent is administered at a dose of about 50 mg/m² to about 100 mg/m². Insome embodiments, the hypomethylating agent is administered at a dose ofabout 75 mg/m². In some embodiments, the hypomethylating agent isadministered once a day. In some embodiments, the hypomethylating agentis administered for 5-7 consecutive days. In some embodiments, thehypomethylating agent is administered for (a) seven consecutive days ondays 1-7 of a 28-day cycle, or (b) five consecutive days on days 1-5,followed by a two-day break, then two consecutive days on days 8-9, of a28-day cycle. In some embodiments, the hypomethylating agent isadministered subcutaneously or intravenously.

In some embodiments, the combination further comprise a CD47 inhibitor,a CD70 inhibitor, a NEDD8 inhibitor, a CDK9 inhibitor, an FLT3inhibitor, a KIT inhibitor, or a p53 activator, or any combinationthereof, e.g., a CD47 inhibitor, a CD70 inhibitor, a NEDD8 inhibitor, aCDK9 inhibitor, an FLT3 inhibitor, a KIT inhibitor, or a p53 activator,all as described herein.

In some embodiments, the myelodysplastic syndrome (MDS) is anintermediate MDS, a high risk MDS, or a very high risk MDS.

In another aspect, the disclosure features a method of treating achronic myelomonocytic leukemia (CMML) in a subject, comprisingadministering to the subject a combination of a TIM-3 inhibitor and ahypomethylating agent.

In some embodiments, the TIM-3 inhibitor comprises an anti-TIM-3antibody molecule. In some embodiments, the TIM-3 inhibitor comprisesMBG453, TSR-022, LY3321367, Sym023, BGB-A425, INCAGN-2390, MBS-986258,RO-7121661, BC-3402, SHR-1702, or LY-3415244. In some embodiments, theTIM-3 inhibitor comprises MBG453. In some embodiments, the TIM-3inhibitor is administered at a dose of about 700 mg to about 900 mg. Insome embodiments, the TIM-3 inhibitor is administered at a dose of about800 mg. In some embodiments, the TIM-3 inhibitor is administered at adose of about 300 mg to about 500 mg. In some embodiments, the TIM-3inhibitor is administered at a dose of about 400 mg. In someembodiments, the TIM-3 inhibitor is administered once every four weeks.In some embodiments, the TIM-3 inhibitor is administered on day 8 of a28-day cycle. In some embodiments, the TIM-3 inhibitor is administeredonce every two weeks. In some embodiments, the TIM-3 inhibitor isadministered at day 8 and day 22 of a 28-day cycle. In some embodiments,the TIM-3 inhibitor is administered once every four weeks. In someembodiments, the TIM-3 inhibitor is administered intravenously. In someembodiments, the TIM-3 inhibitor is administered intravenously over aperiod of about 15 minutes to about 45 minutes. In some embodiments, theTIM-3 inhibitor is administered intravenously over a period of about 30minutes. In some embodiments, the TIM-3 inhibitor is administeredintravenously over a period of about 15 minutes to about 45 minutes. Insome embodiments, the TIM-3 inhibitor is administered intravenously overa period of about 30 minutes.

In some embodiments, the hypomethylating agent comprises azacitidine,decitabine, CC-486 or ASTX727. In some embodiments, the hypomethylatingagent comprises azacitidine. In some embodiments, the hypomethylatingagent is administered at a dose of about 50 mg/m² to about 100 mg/m². Insome embodiments, the hypomethylating agent is administered at a dose ofabout 75 mg/m². In some embodiments, the hypomethylating agent isadministered once a day. In some embodiments, the hypomethylating agentis administered for 5-7 consecutive days. In some embodiments, thehypomethylating agent is administered for (a) seven consecutive days ondays 1-7 of a 28-day cycle, or (b) five consecutive days on days 1-5,followed by a two-day break, then two consecutive days on days 8-9, of a28-day cycle. In some embodiments, the hypomethylating agent (e.g.,azacitidine) is administered subcutaneously or intravenously.

In some embodiments, the combination further comprise a CD47 inhibitor,a CD70 inhibitor, a NEDD8 inhibitor, a CDK9 inhibitor, an FLT3inhibitor, a KIT inhibitor, or a p53 activator, or any combinationthereof, e.g., a CD47 inhibitor, a CD70 inhibitor, a NEDD8 inhibitor, aCDK9 inhibitor, an FLT3 inhibitor, a KIT inhibitor, or a p53 activator,all as described herein.

In some embodiments, the chronic myelomonocytic leukemia (CMML) is aCMML-1 or a CMML-2. In some embodiments, the CMML is a CMML-2.

In another aspect, the disclosure features a combination comprisingMBG453 and azacitidine for use in treating a myelodysplastic syndrome(MDS) in a subject. In some embodiments, MGB453 is administered at adose of 600 mg to 1000 mg (e.g., 800 mg) once every four weeks, andazacitidine is administered at a dose of 50 mg/m² to 100 mg/m² (e.g., 75mg/m²) for (a) seven consecutive days, e.g., on days 1-7 of a 28 daycycle, or (b) five consecutive days, e.g., on days 1-5 of a 28 daycycle, followed by a two day break, then two consecutive days on days 8and 9 of a 28 day cycle. In some embodiments the MDS is intermediateMDS, high risk MDS, or very high risk MDS.

In another aspect, the disclosure features a method of treating amyelodysplastic syndrome (MDS) in a subject comprising administering tothe subject a combination of a MBG453 and azacitidine. In someembodiments, MGB453 is administered at a dose of 600 mg to 1000 mg(e.g., 800 mg) once every four weeks, and azacitidine is administered ata dose of 50 mg/m² to 100 mg/m² (e.g., 75 mg/m²) for (a) sevenconsecutive days, e.g., on days 1-7 of a 28 day cycle, or (b) fiveconsecutive days, e.g., on days 1-5 of a 28 day cycle, followed by a twoday break, then two consecutive days on days 8 and 9 of a 28 day cycle.In some embodiments the MDS is intermediate MDS, high risk MDS, or veryhigh risk MDS.

In another aspect, the disclosure features a combination comprisingMBG453 and azacitidine for use in treating a chronic myelomonocyticleukemia (CMML) in a subject. In some embodiments, MGB453 isadministered at a dose of 600 mg to 1000 mg (e.g., 800 mg) once everyfour weeks, and azacitidine is administered at a dose of 50 mg/m² to 100mg/m² (e.g., 75 mg/m²) for (a) seven consecutive days, e.g., on days 1-7of a 28 day cycle, or (b) five consecutive days, e.g., on days 1-5 of a28 day cycle, followed by a two day break, then two consecutive days ondays 8 and 9 of a 28 day cycle. In some embodiments, the CMML is CMML-2.

In another aspect, the disclosure features a method of treating achronic myelomonocytic leukemia (CMML) in a subject comprisingadministering to the subject a combination of a MBG453 and azacitidine.In some embodiments, MGB453 is administered at a dose of 600 mg to 1000mg (e.g., 800 mg) once every four weeks, and azacitidine is administeredat a dose of 50 mg/m² to 100 mg/m² (e.g., 75 mg/m²) for (a) sevenconsecutive days, e.g., on days 1-7 of a 28 day cycle, or (b) fiveconsecutive days, e.g., on days 1-5 of a 28 day cycle, followed by a twoday break, then two consecutive days on days 8 and 9 of a 28 day cycle.In some embodiments, the CMML is CMML-2.

In another aspect, the disclosure features a method of reducing anactivity (e.g., growth, survival, or viability, or all), of ahematological cancer cell. The method includes contacting the cell witha combination described herein. The method can be performed in asubject, e.g., as part of a therapeutic protocol. The hematologicalcancer cell can be, e.g., a cell from a hematological cancer describedherein, such as a myelodysplastic syndrome (MDS) (e.g., an intermediateMDS, a high risk MDS, or a very high risk MDS) and a chronicmyelomonocytic leukemia (CMML) (e.g., CMML-1 or CMML-2).

In certain embodiments of the methods disclosed herein, the methodfurther includes determining the level of TIM-3 expression in tumorinfiltrating lymphocytes (TILs) in the subject. In other embodiments,the level of TIM-3 expression is determined in a sample (e.g., a liquidbiopsy) acquired from the subject (e.g., using immunohistochemistry). Incertain embodiments, responsive to a detectable level, or an elevatedlevel, of TIM-3 in the subject, the combination is administered. Thedetection steps can also be used, e.g., to monitor the effectiveness ofa therapeutic agent described herein. For example, the detection stepcan be used to monitor the effectiveness of the combination.

In another aspect, the disclosure features a composition (e.g., one ormore compositions or dosage forms), that includes a TIM-3 inhibitor anda hypomethylating agent, as described herein. Formulations, e.g., dosageformulations, and kits, e.g., therapeutic kits, that include a TIM-3inhibitor and a hypomethylating agent, are also described herein. Incertain embodiments, the composition or formulation is used to treat ahematological cancer, e.g., myelodysplastic syndrome (MDS) (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS) and achronic myelomonocytic leukemia (CMML) (e.g., CMML-1 or CMML-2).

Additional features or embodiments of the methods, compositions, dosageformulations, and kits described herein include one or more of thefollowing.

TIM-3 Inhibitors

In some embodiments, the combination described herein comprises a TIM-3inhibitor, e.g., an anti-TIM-3 antibody. In one embodiment, theanti-TIM-3 antibody molecule comprises at least one, two, three, four,five or six complementarity determining regions (CDRs) (or collectivelyall of the CDRs) from a heavy and light chain variable region comprisingan amino acid sequence shown in Table 7 (e.g., from the heavy and lightchain variable region sequences of ABTIM3-hum11 or ABTIM3-hum03disclosed in Table 7), or encoded by a nucleotide sequence shown inTable 7. In some embodiments, the CDRs are according to the Kabatdefinition (e.g., as set out in Table 7). In some embodiments, the CDRsare according to the Chothia definition (e.g., as set out in Table 7).In one embodiment, one or more of the CDRs (or collectively all of theCDRs) have one, two, three, four, five, six or more changes, e.g., aminoacid substitutions (e.g., conservative amino acid substitutions) ordeletions, relative to an amino acid sequence shown in Table 7, orencoded by a nucleotide sequence shown in Table 7.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavychain variable region (VH) comprising a VHCDR1 amino acid sequence ofSEQ ID NO: 801, a VHCDR2 amino acid sequence of SEQ ID NO: 802, and aVHCDR3 amino acid sequence of SEQ ID NO: 803; and a light chain variableregion (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 810, aVLCDR2 amino acid sequence of SEQ ID NO: 811, and a VLCDR3 amino acidsequence of SEQ ID NO: 812, each disclosed in Table 7. In oneembodiment, the anti-TIM-3 antibody molecule comprises a heavy chainvariable region (VH) comprising a VHCDR1 amino acid sequence of SEQ IDNO: 801, a VHCDR2 amino acid sequence of SEQ ID NO: 820, and a VHCDR3amino acid sequence of SEQ ID NO: 803; and a light chain variable region(VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 810, a VLCDR2amino acid sequence of SEQ ID NO: 811, and a VLCDR3 amino acid sequenceof SEQ ID NO: 812, each disclosed in Table 7.

In one embodiment, the anti-TIM-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 806, or an amino acidsequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ IDNO: 806. In one embodiment, the anti-TIM-3 antibody molecule comprises aVL comprising the amino acid sequence of SEQ ID NO: 816, or an aminoacid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQID NO: 816. In one embodiment, the anti-TIM-3 antibody moleculecomprises a VH comprising the amino acid sequence of SEQ ID NO: 822, oran amino acid sequence at least 85%, 90%, 95%, or 99% identical orhigher to SEQ ID NO: 822. In one embodiment, the anti-TIM-3 antibodymolecule comprises a VL comprising the amino acid sequence of SEQ ID NO:826, or an amino acid sequence at least 85%, 90%, 95%, or 99% identicalor higher to SEQ ID NO: 826. In one embodiment, the anti-TIM-3 antibodymolecule comprises a VH comprising the amino acid sequence of SEQ ID NO:806 and a VL comprising the amino acid sequence of SEQ ID NO: 816. Inone embodiment, the anti-TIM-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 822 and a VL comprisingthe amino acid sequence of SEQ ID NO: 826.

In one embodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 807, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 807. In oneembodiment, the antibody molecule comprises a VL encoded by thenucleotide sequence of SEQ ID NO: 817, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 817. In oneembodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 823, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 823. In oneembodiment, the antibody molecule comprises a VL encoded by thenucleotide sequence of SEQ ID NO: 827, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 827. In oneembodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 807 and a VL encoded by the nucleotidesequence of SEQ ID NO: 817. In one embodiment, the antibody moleculecomprises a VH encoded by the nucleotide sequence of SEQ ID NO: 823 anda VL encoded by the nucleotide sequence of SEQ ID NO: 827.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavychain comprising the amino acid sequence of SEQ ID NO: 808, or an aminoacid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQID NO: 808. In one embodiment, the anti-TIM-3 antibody moleculecomprises a light chain comprising the amino acid sequence of SEQ ID NO:818, or an amino acid sequence at least 85%, 90%, 95%, or 99% identicalor higher to SEQ ID NO: 818. In one embodiment, the anti-TIM-3 antibodymolecule comprises a heavy chain comprising the amino acid sequence ofSEQ ID NO: 824, or an amino acid sequence at least 85%, 90%, 95%, or 99%identical or higher to SEQ ID NO: 824. In one embodiment, the anti-TIM-3antibody molecule comprises a light chain comprising the amino acidsequence of SEQ ID NO: 828, or an amino acid sequence at least 85%, 90%,95%, or 99% identical or higher to SEQ ID NO: 828. In one embodiment,the anti-TIM-3 antibody molecule comprises a heavy chain comprising theamino acid sequence of SEQ ID NO: 808 and a light chain comprising theamino acid sequence of SEQ ID NO: 818. In one embodiment, the anti-TIM-3antibody molecule comprises a heavy chain comprising the amino acidsequence of SEQ ID NO: 824 and a light chain comprising the amino acidsequence of SEQ ID NO: 828.

In one embodiment, the antibody molecule comprises a heavy chain encodedby the nucleotide sequence of SEQ ID NO: 809, or a nucleotide sequenceat least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 809. Inone embodiment, the antibody molecule comprises a light chain encoded bythe nucleotide sequence of SEQ ID NO: 819, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 819. Inone embodiment, the antibody molecule comprises a heavy chain encoded bythe nucleotide sequence of SEQ ID NO: 825, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 825. Inone embodiment, the antibody molecule comprises a light chain encoded bythe nucleotide sequence of SEQ ID NO: 829, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 829. Inone embodiment, the antibody molecule comprises a heavy chain encoded bythe nucleotide sequence of SEQ ID NO: 809 and a light chain encoded bythe nucleotide sequence of SEQ ID NO: 819. In one embodiment, theantibody molecule comprises a heavy chain encoded by the nucleotidesequence of SEQ ID NO: 825 and a light chain encoded by the nucleotidesequence of SEQ ID NO: 829.

In some embodiments, the anti-TIM-3 antibody is MBG453, which isdisclosed in WO2015/117002. MBG453 is also sometimes referred to assabatolimab herein.

Other Exemplary TIM-3 Inhibitors

In one embodiment, the anti-TIM-3 antibody molecule is TSR-022(AnaptysBio/Tesaro). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of TSR-022. In oneembodiment, the anti-TIM-3 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain variable region sequence and/or light chain variable regionsequence, or the heavy chain sequence and/or light chain sequence ofAPE5137 or APE5121, e.g., as disclosed in Table 8. APE5137, APE5121, andother anti-TIM-3 antibodies are disclosed in WO 2016/161270,incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule is the antibodyclone F38-2E2. In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of F38-2E2.

In one embodiment, the anti-TIM-3 antibody molecule is LY3321367 (EliLilly). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of LY3321367.

In one embodiment, the anti-TIM-3 antibody molecule is Sym023(Symphogen). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of Sym023.

In one embodiment, the anti-TIM-3 antibody molecule is BGB-A425(Beigene). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of BGB-A425.

In one embodiment, the anti-TIM-3 antibody molecule is INCAGN-2390(Agenus/Incyte). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain or light chainsequence of INCAGN-2390.

In one embodiment, the anti-TIM-3 antibody molecule is MBS-986258(BMS/Five Prime). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of MBS-986258.

In one embodiment, the anti-TIM-3 antibody molecule is RO-7121661(Roche). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of RO-7121661.

In one embodiment, the anti-TIM-3 antibody molecule is LY-3415244 (EliLilly). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of LY-3415244.

Further known anti-TIM-3 antibodies include those described, e.g., in WO2016/111947, WO 2016/071448, WO 2016/144803, U.S. Pat. Nos. 8,552,156,8,841,418, and 9,163,087, incorporated by reference in their entirety.

In one embodiment, the anti-TIM-3 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on TIM-3 as, one ofthe anti-TIM-3 antibodies described herein.

In one embodiment, the anti-TIM-3 antibody molecule is BC-3402 (WuxiZhikanghongyi Biotechnology). In one embodiment, the anti-TIM-3 antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain variable region sequence and/orlight chain variable region sequence, or the heavy chain sequence and/orlight chain sequence of BC-3402.

In one embodiment, the anti-TIM-3 antibody molecule is SHR-1702(Medicine Co Ltd.). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of SHR-1702. SHR-1702 is disclosed, e.g., in WO2020/038355.

Hypomethylating Agents

In some embodiments, the combination described herein comprises ahypomethylating agent. In some embodiments, the hypomethylating agent isused in combination with a TIM-3 inhibitor (e.g., an anti-TIM-3 antibodymolecule). In some embodiments, the hypomethylating agent is used incombination with a TIM-3 inhibitor (e.g., an anti-TIM-3 antibodymolecule) to treat a hematological cancer. In some embodiments, thehematological cancer is a myelodysplastic syndrome (MDS) (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS) and achronic myelomonocytic leukemia (CMML) (e.g., CMML-1 or CMML-2). In someembodiments, the hypomethylating agent is azacitidine, decitabine,CC-486 or ASTX727. In some embodiments, the hypomethylating agent isazacitidine. In certain embodiments, the hypomethylating agent (e.g.,azacitidine) is used in combination with an anti-TIM-3 antibody molecule(e.g., MBG453) to treat an MDS. In certain embodiments, thehypomethylating agent (e.g., azacitidine) is used in combination with ananti-TIM-3 antibody molecule (e.g., MBG453) to treat a CMML, e.g., aCMML-2. In certain embodiments, at least five (e.g., 5, 6, 7, 8, 9, 10,or more) doses of the hypomethylating agent (e.g., azacitidine) areadministered in a dosing cycle prior to administration of the first doseof the anti-TIM-3 antibody molecule (e.g., MBG453). In certainembodiments, the anti-TIM-3 antibody molecule (e.g., MBG453) and thehypomethylating agent (e.g., azacitidine) are administered on the sameday, e.g., day 8 of a 28-day cycle. In certain embodiments, thehypomethylating agent is administered prior to the anti-TIM-3 antibodymolecule (e.g., MBG453), e.g., at least 30 minutes prior toadministration of the anti-TIM-3 antibody molecule (e.g., MBG453).

Therapeutic Use

Without wishing to be bound by theory, it is believed that in someembodiments, the combinations described herein can inhibit, reduce, orneutralize one or more activities of TIM-3, or DNA methyltransferase,resulting in, e.g., one or more of immune checkpoint inhibition,hypomethylation, or cytotoxicity. Thus, the combinations describedherein can be used to treat or prevent disorders (e.g., cancer), whereenhancing an immune response in a subject is desired.

Accordingly, in another aspect, a method of modulating an immuneresponse in a subject is provided. The method comprises administering tothe subject a therapeutically effective amount of a combinationdescribed herein, e.g., in accordance with a dosage regimen describedherein, such that the immune response in the subject is modulated. Inone embodiment, the combination enhances, stimulates or increases theimmune response in the subject. The subject can be a mammal, e.g., aprimate, preferably a higher primate, e.g., a human (e.g., a patienthaving, or at risk of having, a disorder described herein). In oneembodiment, the subject is in need of enhancing an immune response. Inone embodiment, the subject has, or is at risk of, having a disorderdescribed herein, e.g., a cancer as described herein. In certainembodiments, the subject is, or is at risk of being, immunocompromised.For example, the subject is undergoing or has undergone achemotherapeutic treatment and/or radiation therapy. Alternatively, orin combination, the subject is, or is at risk of being,immunocompromised as a result of an infection. In certain embodiments,the subject is unfit for a chemotherapy, e.g., an intensive inductionchemotherapy.

In one aspect, a method of treating (e.g., one or more of reducing,inhibiting, or delaying progression) a cancer in a subject is provided.The method comprises administering to the subject a therapeuticallyeffective amount of a combination disclosed herein, e.g., in accordancewith a dosage regimen described herein, thereby treating the cancer inthe subject.

In certain embodiments, the cancer treated with the combinationincludes, but is not limited to, a hematological cancer (e.g., leukemia,lymphoma, or myeloma), a solid tumor, and a metastatic lesion. In oneembodiment, the cancer a hematological cancer. Examples of hematologicalcancers include, e.g., a leukemia (e.g., an acute myeloid leukemia (AML)or A chronic lymphocytic leukemia (CLL), a lymphoma (e.g., smalllymphocytic lymphoma (SLL)), and a myeloma (e.g., a multiple myeloma(MM)). The cancer may be at an early, intermediate, late stage ormetastatic cancer.

In certain embodiments, the hematological cancer treated with thecombination includes, but is not limited to, myelodysplastic syndrome(MDS) (e.g., an intermediate MDS, a high risk MDS, or a very high riskMDS) or a chronic myelomonocytic leukemia (CMML) (e.g., a CMML-1 or aCMML-2). In certain embodiments, the cancer treated with the combinationis a CMML-2.

In certain embodiments, the cancer is an MSI-high cancer. In someembodiments, the cancer is a metastatic cancer. In other embodiments,the cancer is an advanced cancer. In other embodiments, the cancer is arelapsed or refractory cancer.

In other embodiments, the subject has, or is identified as having, TIM-3expression in tumor-infiltrating lymphocytes (TILs). In one embodiment,the cancer microenvironment has an elevated level of TIM-3 expression.In one embodiment, the cancer microenvironment has an elevated level ofPD-L1 expression. Alternatively, or in combination, the cancermicroenvironment can have increased IFN□ and/or CD8 expression.

In some embodiments, the subject has, or is identified as having, atumor that has one or more of high PD-L1 level or expression, or asbeing tumor infiltrating lymphocyte (TIL)+ (e.g., as having an increasednumber of TILs), or both. In certain embodiments, the subject has, or isidentified as having, a tumor that has high PD-L1 level or expressionand that is TIL+. In some embodiments, the methods described hereinfurther include identifying a subject based on having a tumor that hasone or more of high PD-L1 level or expression, or as being TIL+, orboth. In certain embodiments, the methods described herein furtherinclude identifying a subject based on having a tumor that has highPD-L1 level or expression and as being TIL+. In some embodiments, tumorsthat are TIL+ are positive for CD8 and IFNγ. In some embodiments, thesubject has, or is identified as having, a high percentage of cells thatare positive for one, two or more of PD-L1, CD8, and/or IFNγ. In certainembodiments, the subject has or is identified as having a highpercentage of cells that are positive for all of PD-L1, CD8, and IFNγ.

In some embodiments, the methods described herein further includeidentifying a subject based on having a high percentage of cells thatare positive for one, two or more of PD-L1, CD8, and/or IFNγ. In certainembodiments, the methods described herein further include identifying asubject based on having a high percentage of cells that are positive forall of PD-L1, CD8, and IFNγ. In some embodiments, the subject has, or isidentified as having, one, two or more of PD-L1, CD8, and/or IFNγ, andone or more of a hematological cancer, e.g., a leukemia (e.g., an AML orCLL), a lymphoma, (e.g., an SLL), and/or a myeloma (e.g., an MM). Incertain embodiments, the methods described herein further describeidentifying a subject based on having one, two or more of PD-L1, CD8,and/or IFNγ, and one or more of a leukemia (e.g., an AML or CLL), alymphoma, (e.g., an SLL), and/or a myeloma (e.g., an MM).

Methods, compositions, and formulations disclosed herein are useful fortreating metastatic lesions associated with the aforementioned cancers.

Still further, the invention provides a method of enhancing an immuneresponse to an antigen in a subject, comprising administering to thesubject: (i) the antigen; and (ii) a combination described herein, inaccordance with a dosage regimen described herein, such that an immuneresponse to the antigen in the subject is enhanced. The antigen can be,for example, a tumor antigen, a viral antigen, a bacterial antigen or anantigen from a pathogen.

The combination described herein can be administered to the subjectsystemically (e.g., orally, parenterally, subcutaneously, intravenously,rectally, intramuscularly, intraperitoneally, intranasally,transdermally, or by inhalation or intracavitary installation),topically, or by application to mucous membranes, such as the nose,throat and bronchial tubes. In certain embodiments, the anti-TIM-3antibody molecule is administered intravenously at a flat dose describedherein.

Immunomodulators

The combinations described herein (e.g., a combination comprising atherapeutically effective amount of an anti-TIM-3 antibody moleculedescribed herein) can be used further in combination with one or moreimmunomodulators.

In certain embodiments, the immunomodulator is an inhibitor of an immunecheckpoint molecule. In one embodiment, the immunomodulator is aninhibitor of PD-1, PD-L1, PD-L2, CTLA-4, LAG-3, CEACAM (e.g., CEACAM-1,-3 and/or -5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF beta. Inone embodiment, the inhibitor of an immune checkpoint molecule inhibitsPD-1, PD-L1, LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), CTLA-4, orany combination thereof.

Inhibition of an inhibitory molecule can be performed at the DNA, RNA orprotein level. In embodiments, an inhibitory nucleic acid (e.g., adsRNA, siRNA or shRNA), can be used to inhibit expression of aninhibitory molecule. In other embodiments, the inhibitor of aninhibitory signal is, a polypeptide e.g., a soluble ligand (e.g.,PD-1-Ig or CTLA-4 Ig), or an antibody molecule that binds to theinhibitory molecule; e.g., an antibody molecule that binds to PD-1,PD-L1, PD-L2, CEACAM (e.g., CEACAM-1, -3 and/or -5), CTLA-4, LAG-3,VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF beta, or a combinationthereof.

In certain embodiments, the combination comprises an anti-TIM-3 antibodymolecule that is in the form of a bispecific or multispecific antibodymolecule. In one embodiment, the bispecific antibody molecule has afirst binding specificity to TIM-3 and a second binding specificity,e.g., a second binding specificity to, PD-1, PD-L1, CEACAM (e.g.,CEACAM-1, -3 and/or -5), LAG-3, or PD-L2. In one embodiment, thebispecific antibody molecule binds to (i) PD-1 or PD-L1 (ii) and TIM-3.In another embodiment, the bispecific antibody molecule binds to TIM-3and LAG-3. In another embodiment, the bispecific antibody molecule bindsto TIM-3 and CEACAM (e.g., CEACAM-1, -3 and/or -5). In anotherembodiment, the bispecific antibody molecule binds to TIM-3 andCEACAM-1. In still another embodiment, the bispecific antibody moleculebinds to TIM-3 and CEACAM-3. In yet another embodiment, the bispecificantibody molecule binds to TIM-3 and CEACAM-5.

In other embodiments, the combination further comprises a bispecific ormultispecific antibody molecule. In another embodiment, the bispecificantibody molecule binds to PD-1 or PD-L1. In yet another embodiment, thebispecific antibody molecule binds to PD-1 and PD-L2. In anotherembodiment, the bispecific antibody molecule binds to CEACAM (e.g.,CEACAM-1, -3 and/or -5) and LAG-3.

Any combination of the aforesaid molecules can be made in amultispecific antibody molecule, e.g., a trispecific antibody thatincludes a first binding specificity to TIM-3, and a second and thirdbinding specificities to two or more of: PD-1, PD-L1, CEACAM (e.g.,CEACAM-1, -3 and/or -5), LAG-3, or PD-L2.

In certain embodiments, the immunomodulator is an inhibitor of PD-1,e.g., human PD-1. In another embodiment, the immunomodulator is aninhibitor of PD-L1, e.g., human PD-L1. In one embodiment, the inhibitorof PD-1 or PD-L1 is an antibody molecule to PD-1 or PD-L1 (e.g., ananti-PD-1 or anti-PD-L1 antibody molecule as described herein).

The combination of the PD-1 or PD-L1 inhibitor with the anti-TIM-3antibody molecule can further include one or more additionalimmunomodulators, e.g., in combination with an inhibitor of LAG-3,CEACAM (e.g., CEACAM-1, -3 and/or -5) or CTLA-4. In one embodiment, theinhibitor of PD-1 or PD-L1 (e.g., the anti-PD-1 or PD-L1 antibodymolecule) is administered in combination with the anti-TIM-3 antibodymolecule and a LAG-3 inhibitor (e.g., an anti-LAG-3 antibody molecule).In another embodiment, the inhibitor of PD-1 or PD-L1 (e.g., theanti-PD-1 or PD-L1 antibody molecule) is administered in combinationwith the anti-TIM-3 antibody molecule and a CEACAM inhibitor (e.g.,CEACAM-1, -3 and/or -5 inhibitor), e.g., an anti-CEACAM antibodymolecule. In another embodiment, the inhibitor of PD-1 or PD-L1 (e.g.,the anti-PD-1 or PD-L1 antibody molecule) is administered in combinationwith the anti-TIM-3 antibody molecule and a CEACAM-1 inhibitor (e.g., ananti-CEACAM-1 antibody molecule). In another embodiment, the inhibitorof PD-1 or PD-L1 (e.g., the anti-PD-1 or PD-L1 antibody molecule) isadministered in combination with the anti-TIM-3 antibody molecule and aCEACAM-5 inhibitor (e.g., an anti-CEACAM-5 antibody molecule). In yetother embodiments, the inhibitor of PD-1 or PD-L1 (e.g., the anti-PD-1or PD-L1 antibody molecule) is administered in combination with theanti-TIM-3 antibody molecule, a LAG-3 inhibitor (e.g., an anti-LAG-3antibody molecule), and a TIM-3 inhibitor (e.g., an anti-TIM-3 antibodymolecule). Other combinations of immunomodulators with the anti-TIM-3antibody molecule and a PD-1 inhibitor (e.g., one or more of PD-L2,CTLA-4, LAG-3, CEACAM (e.g., CEACAM-1, -3 and/or -5), VISTA, BTLA,TIGIT, LAIR1, CD160, 2B4 and/or TGF beta) are also within the presentinvention. Any of the antibody molecules known in the art or disclosedherein can be used in the aforesaid combinations of inhibitors ofcheckpoint molecule.

In other embodiments, the immunomodulator is an inhibitor of CEACAM(e.g., CEACAM-1, -3 and/or -5), e.g., human CEACAM (e.g., CEACAM-1, -3and/or -5). In one embodiment, the immunomodulator is an inhibitor ofCEACAM-1, e.g., human CEACAM-1. In another embodiment, theimmunomodulator is an inhibitor of CEACAM-3, e.g., human CEACAM-3. Inanother embodiment, the immunomodulator is an inhibitor of CEACAM-5,e.g., human CEACAM-5. In one embodiment, the inhibitor of CEACAM (e.g.,CEACAM-1, -3 and/or -5) is an antibody molecule to CEACAM (e.g.,CEACAM-1, -3 and/or -5). The combination of the CEACAM (e.g., CEACAM-1,-3 and/or -5) inhibitor and the anti-TIM-3 antibody molecule can furtherinclude one or more additional immunomodulators, e.g., in combinationwith an inhibitor of LAG-3, PD-1, PD-L1 or CTLA-4.

In other embodiments, the immunomodulator is an inhibitor of LAG-3,e.g., human LAG-3. In one embodiment, the inhibitor of LAG-3 is anantibody molecule to LAG-3. The combination of the LAG-3 inhibitor andthe anti-TIM-3 antibody molecule can further include one or moreadditional immunomodulators, e.g., in combination with an inhibitor ofCEACAM (e.g., CEACAM-1, -3 and/or -5), PD-1, PD-L1 or CTLA-4.

In certain embodiments, the immunomodulator used in the combinationsdisclosed herein (e.g., in combination with a therapeutic agent chosenfrom an antigen-presentation combination) is an activator or agonist ofa costimulatory molecule. In one embodiment, the agonist of thecostimulatory molecule is chosen from an agonist (e.g., an agonisticantibody or antigen-binding fragment thereof, or a soluble fusion) ofOX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278),4-1BB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7,NKp80, CD160, B7-H3, or CD83 ligand.

In other embodiments, the immunomodulator is a GITR agonist. In oneembodiment, the GITR agonist is an antibody molecule to GITR. Theanti-GITR antibody molecule and the anti-TIM-3 antibody molecule may bein the form of separate antibody composition, or as a bispecificantibody molecule. The combination of the GITR agonist with theanti-TIM-3 antibody molecule can further include one or more additionalimmunomodulators, e.g., in combination with an inhibitor of PD-1, PD-L1,CTLA-4, CEACAM (e.g., CEACAM-1, -3 and/or -5), or LAG-3. In someembodiments, the anti-GITR antibody molecule is a bispecific antibodythat binds to GITR and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3and/or -5), or LAG-3. In other embodiments, a GITR agonist can beadministered in combination with one or more additional activators ofcostimulatory molecules, e.g., an agonist of OX40, CD2, CD27, CD28, CDS,ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137), CD30, CD40,BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, or CD83ligand.

In other embodiments, the immunomodulator is an OX40 agonist. In oneembodiment, the OX40 agonist is an antibody molecule to OX40. The OX40antibody molecule and the anti-TIM-3 antibody molecule may be in theform of separate antibody composition, or as a bispecific antibodymolecule. The combination of the OX40 agonist with the anti-TIM-3antibody molecule can further include one or more additionalimmunomodulators, e.g., in combination with an inhibitor of PD-1, PD-L1,CTLA-4, CEACAM (e.g., CEACAM-1, -3 and/or -5), or LAG-3. In someembodiments, the anti-OX40 antibody molecule is a bispecific antibodythat binds to OX40 and PD-1, PD-L1, CTLA-4, CEACAM (e.g., CEACAM-1, -3and/or -5), or LAG-3. In other embodiments, the OX40 agonist can beadministered in combination with other costimulatory molecule, e.g., anagonist of GITR, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS(CD278), 4-1BB (CD137), CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C,SLAMF7, NKp80, CD160, B7-H3, or CD83 ligand.

It is noted that only exemplary combinations of inhibitors of checkpointinhibitors or agonists of costimulatory molecules are provided herein.Additional combinations of these agents are within the scope of thepresent invention.

Biomarkers

In certain embodiments, any of the methods or use disclosed hereinfurther includes evaluating or monitoring the effectiveness of a therapy(e.g., a combination therapy) described herein, in a subject (e.g., asubject having a cancer, e.g., a cancer described herein). The methodincludes acquiring a value of effectiveness to the therapy, wherein saidvalue is indicative of the effectiveness of the therapy.

In embodiments, the value of effectiveness to the therapy comprises ameasure of one, two, three, four, five, six, seven, eight, nine or more(e.g., all) of the following:

(i) a parameter of a tumor infiltrating lymphocyte (TIL) phenotype;

(ii) a parameter of a myeloid cell population;

(iii) a parameter of a surface expression marker;

(iv) a parameter of a biomarker of an immunologic response;

(v) a parameter of a systemic cytokine modulation;

(vi) a parameter of circulating free DNA (cfDNA);

(vii) a parameter of systemic immune-modulation;

(viii) a parameter of microbiome;

(ix) a parameter of a marker of activation in a circulating immune cell;

(x) a parameter of a circulating cytokine; or

(xi) a parameter of minimal residual disease (MRD)

In some embodiments, the parameter of a TIL phenotype comprises thelevel or activity of one, two, three, four or more (e.g., all) ofHematoxylin and eosin (H&E) staining for TIL counts, CD8, FOXP3, CD4, orCD3, in the subject, e.g., in a sample from the subject (e.g., a tumorsample).

In some embodiments, the parameter of a myeloid cell populationcomprises the level or activity of one or both of CD68 or CD163, in thesubject, e.g., in a sample from the subject (e.g., a tumor sample).

In some embodiments, the parameter of a surface expression markercomprises the level or activity of one, two, three or more (e.g., all)of TIM-3, PD-1, PD-L1, or LAG-3, in the subject, e.g., in a sample fromthe subject (e.g., a tumor sample). In certain embodiments, the level ofTIM-3, PD-1, PD-L1, or LAG-3 is determined by immunohistochemistry(IHC). In certain embodiments, the level of TIM-3 is determined.

In some embodiments, the parameter of a biomarker of an immunologicresponse comprises the level or sequence of one or more nucleicacid-based markers, in the subject, e.g., in a sample from the subject(e.g., a tumor sample).

In some embodiments, the parameter of systemic cytokine modulationcomprises the level or activity of one, two, three, four, five, six,seven, eight, or more (e.g., all) of IL-18, IFN-γ, ITAC (CXCL11), IL-6,IL-10, IL-4, IL-17, IL-15, or TGF-beta, in the subject, e.g., in asample from the subject (e.g., a blood sample, e.g., a plasma sample).

In some embodiments, the parameter of cfDNA comprises the sequence orlevel of one or more circulating tumor DNA (cfDNA) molecules, in thesubject, e.g., in a sample from the subject (e.g., a blood sample, e.g.,a plasma sample).

In some embodiments, the parameter of systemic immune-modulationcomprises phenotypic characterization of an activated immune cell, e.g.,a CD3-expressing cell, a CD8-expressing cell, or both, in the subject,e.g., in a sample from the subject (e.g., a blood sample, e.g., a PBMCsample).

In some embodiments, the parameter of microbiome comprises the sequenceor expression level of one or more genes in the microbiome, in thesubject, e.g., in a sample from the subject (e.g., a stool sample).

In some embodiments, the parameter of a marker of activation in acirculating immune cell comprises the level or activity of one, two,three, four, five or more (e.g., all) of circulating CD8+, HLA-DR+Ki67+,T cells, IFN-γ, IL-18, or CXCL11 (IFN-γ induced CCK) expressing cells,in a sample (e.g., a blood sample, e.g., a plasma sample).

In some embodiments, the parameter of a circulating cytokine comprisesthe level or activity of IL-6, in the subject, e.g., in a sample fromthe subject (e.g., a blood sample, e.g., a plasma sample).

In some embodiments, the parameter of minimal residual disease comprisesa measurement of soluble biomarkers, e.g., soluble TIM-3 and/or anMDS-related gene, e.g., DNMT3, ASXL1, TET2, RUNX1, TP53, or anycombination thereof, in the subject, e.g., in a sample from the subject(e.g., a bone marrow sample, or blood sample, e.g., a plasma sample). Insome embodiments, the minimal residual disease (MRD) parameter ismeasured using cellular (e.g., Multiparameter Flow Cytometry (MFC))and/or molecular (e.g. Next Generation Sequencing (NGS)) methods (seeJongen-Lavrencic M, Grob T, Hanekamp D, et al (2018) Molecular MinimalResidual Disease in Acute Myeloid Leukemia. N Engl J Med; 378(13):1189-99).

In some embodiments of any of the methods disclosed herein, the therapycomprises a combination of an anti-TIM-3 antibody molecule describedherein and a second inhibitor of an immune checkpoint molecule, e.g., aninhibitor of PD-1 (e.g., an anti-PD-1 antibody molecule) or an inhibitorof PD-L1 (e.g., an anti-PD-L1 antibody molecule).

In some embodiments of any of the methods disclosed herein, the measureof one or more of (i)-(xi) is obtained from a sample acquired from thesubject. In some embodiments, the sample is chosen from a tumor sample,a blood sample (e.g., a plasma sample or a PBMC sample), or a stoolsample.

In some embodiments of any of the methods disclosed herein, the subjectis evaluated prior to receiving, during, or after receiving, thetherapy.

In some embodiments of any of the methods disclosed herein, the measureof one or more of (i)-(xi) evaluates a profile for one or more of geneexpression, flow cytometry or protein expression.

In some embodiments of any of the methods disclosed herein, the presenceof an increased level or activity of one, two, three, four, five, ormore (e.g., all) of circulating CD8+, HLA-DR+Ki67+, T cells, IFN-γ,IL-18, or CXCL11 (IFN-γ induced CCK) expressing cells, and/or thepresence of an decreased level or activity of IL-6, in the subject orsample, is a positive predictor of the effectiveness of the therapy.

Alternatively, or in combination with the methods disclosed herein,responsive to said value, performing one, two, three, four or more(e.g., all) of:

(i) administering to the subject the therapy;

(ii) administered an altered dosing of the therapy;

(iii) altering the schedule or time course of the therapy;

(iv) administering to the subject an additional agent (e.g., atherapeutic agent described herein) in combination with the therapy; or

(v) administering to the subject an alternative therapy.

Additional Embodiments

In certain embodiments, any of the methods disclosed herein furtherincludes identifying in a subject or a sample (e.g., a subject's samplecomprising cancer cells and/or immune cells such as TILs) the presenceof TIM-3, thereby providing a value for TIM-3. The method can furtherinclude comparing the TIM-3 value to a reference value, e.g., a controlvalue. If the TIM-3 value is greater than the reference value, e.g., thecontrol value, administering a therapeutically effective amount of thecombination described herein that comprises an anti-TIM-3 antibodymolecule described herein to the subject, and optionally, in combinationwith a second therapeutic agent (e.g., a hypomethylating agent, e.g.,azacitidine), or a procedure, or modality described herein, therebytreating a cancer.

In other embodiments, any of the methods disclosed herein furtherincludes identifying in a subject or a sample (e.g., a subject's samplecomprising cancer cells and/or immune cells such as TILs) the presenceof PD-L1, thereby providing a value for PD-L1. The method can furtherinclude comparing the PD-L1 value to a reference value, e.g., a controlvalue. If the PD-L1 value is greater than the reference value, e.g., thecontrol value, administering a therapeutically effective amount of ananti-TIM-3 antibody molecule described herein to the subject, andoptionally, in combination with a second therapeutic agent, procedure,or modality described herein, thereby treating a cancer.

In other embodiments, any of the methods disclosed herein furtherincludes identifying in a subject or a sample (e.g., a subject's samplecomprising cancer cells and optionally immune cells such as TILs) thepresence of one, two or all of PD-L1, CD8, or IFN-γ, thereby providing avalue for one, two or all of PD-L1, CD8, and IFN-γ. The method canfurther include comparing the PD-L1, CD8, and/or IFN-γ values to areference value, e.g., a control value. If the PD-L1, CD8, and/or IFN-γvalues are greater than the reference value, e.g., the control values,administering a therapeutically effective amount of an anti-TIM-3antibody molecule described herein to the subject, and optionally, incombination with a second therapeutic agent, procedure, or modalitydescribed herein, thereby treating a cancer.

The subject may have a cancer described herein, such as a hematologicalcancer or a solid tumor, e.g., a leukemia (e.g., an acute myeloidleukemia (AML), e.g., a relapsed or refractory AML or a de novo AML), alymphoma, a myeloma, an ovarian cancer, a lung cancer (e.g., a smallcell lung cancer (SCLC) or a non-small cell lung cancer (NSCLC)), amesothelioma, a skin cancer (e.g., a Merkel cell carcinoma (MCC) or amelanoma), a kidney cancer (e.g., a renal cell carcinoma), a bladdercancer, a soft tissue sarcoma (e.g., a hemangiopericytoma (HPC)), a bonecancer (e.g., a bone sarcoma), a colorectal cancer, a pancreatic cancer,a nasopharyngeal cancer, a breast cancer, a duodenal cancer, anendometrial cancer, an adenocarcinoma (an unknown adenocarcinoma), aliver cancer (e.g., a hepatocellular carcinoma), a cholangiocarcinoma, asarcoma. The subject may have a myelodysplastic syndrome (MDS), e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS. The subjectmay have a chronic myelomonocytic leukemia (CMML), e.g., a CMML-1 or aCMML-2.

In certain embodiments, the combination disclosed herein results in alevel of minimal residual disease (MRD) less than 1%, 0.5%, 0.2%, 0.1%,0.05%, 0.02%, or 0.01%, in the subject. In other embodiments, thecombination disclosed herein results in a level of MRD in the subjectthat is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, 500,or 1000-fold lower, compared to a reference MRD level, e.g., the levelof MRD in the subject before receiving the combination. In otherembodiments, the subject described herein has, or is identified ashaving, a level of MRD less than 1%, 0.5%, 0.2%, 0.1%, 0.05%, 0.02%, or0.01%, after receiving the combination. In other embodiments, thesubject disclosed herein has, or is identified as having, a level of MRDthat is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, or 100, 200,500, or 1000-fold lower, compared to a reference MRD level, e.g., thelevel of MRD before receiving the combination. In other embodiments, anyof the methods disclosed herein further comprises determining the levelof MRD in a sample from the subject. In other embodiments, thecombination disclosed herein further comprises determining the durationof remission in the subject.

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety.

Other features, objects, and advantages of the invention will beapparent from the description and drawings, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph depicting the impact of MBG453 on the interactionbetween TIM-3 and galectin-9. Competition was assessed as a measure ofthe ability of the antibody to block Gal9-SULFOTag signal to TIM-3receptor, which is shown on the Y-axis. Concentration of the antibody isshown on the X-axis.

FIG. 2 is graph showing that MBG453 mediates modest antibody-dependentcellular phagocytosis (ADCP). The percentage of phagocytosis wasquantified at various concentrations tested of MBG453, Rituximab, and acontrol hIgG4 monoclonal antibody (mAB).

FIG. 3 is a graph demonstrating MBG453 engagement of FcγR1a as measuredby luciferase activity. The activation of the NFAT dependent reportergene expression induced by the binding of MBG453 or the anti-CD20MabThera reference control to FcγR1a was quantified by luciferaseactivity at various concentrations of the antibody tested.

FIG. 4 shows that MBG453 enhances immune-mediated killing of decitabinepre-treated AML cells.

FIG. 5 is a graph depicting the anti-leukemic activity of MBG453 withand without decitabine in the AML patient-derived xenograft (PDX) model,HAMLX21432. MBG453 was administered i.p. at 10 mg/kg, once weekly(starting at day 6 of dosing) either as a single agent or in combinationwith decitabine i.p. at 1 mg/kg, once daily for a total of 5 doses (frominitiation of dosing). Initial group size: 4 animals Body weights wererecorded weekly during a 21-day dosing period that commenced on day 27post implantation (AML PDX model #21432 2×10⁶ cells/animal) All finaldata were recorded on day 56. Leukemic burden was measured as apercentage of human CD45+ cells in peripheral blood by FACS analysis.

FIG. 6 is a graph depicting the anti-leukemic activity of MBG453 withand without decitabine in the AML patient-derived xenograft (PDX) model,HAMLX5343. Treatments started on day 32 post implantation (2 millioncells/animal) MBG453 was administered i.p. at 10 mg/kg, once weekly(starting on day 6 of dosing), either as a single agent or incombination with decitabine i.p. at 1 mg/kg, once daily for a total of 5doses (from initiation of dosing). Initial group size: 4 animals Bodyweights were recorded weekly during a 21 day dosing period. All finaldata were recorded on day 56. Leukemic burden was measured as apercentage of CD45+ cells in peripheral blood by FACS analysis.

FIG. 7 is a graph depicting MBG453 enhanced killing of THP-1 AML cellsthat were engineered to overexpress TIM-3 relative to parental controlTHP-1 cells. The ratio between TIM-3-expressing THP-1 cells and parentalTHP-1 cells (“fold” in y-axis of graph) was calculated and normalized toconditions without anti-CD3/anti-CD28 bead stimulation. The x-axis ofthe graph denotes the stimulation amount as number of beads per cell.Data represents one of two independent experiments.

DETAILED DESCRIPTION

T-cell immunoglobulin and mucin domain-containing 3 (TIM-3; also knownas hepatitis A virus cellular receptor 2) is a negative regulator of Tcells. TIM-3 was initially described as an inhibitory protein expressedon activated T helper (Th) 1 CD4+ and cytotoxic CD8+ T cells thatsecrete interferon-gamma (IFN-γ) (Monney et al. Nature. 2002;415(6871):536-541; Sánchez Fueyo et al. Nat Immunol. 2003;4(11):1093-101). TIM-3 is enriched on FoxP3+ Tregs and constitutivelyexpressed on DCs, monocytes/macrophages, and NK cells (Anderson et al.Science. 2007; 318(5853):1141-1143; Ndhlovu et al. Blood. 2012; 119(16):3734-3743). Patients with myelodysplastic syndrome (MDS) overexpressTIM-3, which inhibits immune recognition by cytotoxic T cells (Kikushigeet al. Cell Stem Cell. 2010; 7(6): 708-717), and TIM-3 expression levelson MDS blasts increases as MDS progresses to the advanced stage. It hasbeen observed that the proliferation of TIM-3 and MDS blasts isinhibited by the blockade of TIM-3 using an anti-TIM-3 antibody (Asayamaet al. Oncotarget 2017; 8(51):88904-17). Additional preclinical andclinical anti-cancer activities have been reported for TIM-3 blockade(Kikushige et al. Cell Stem Cell. 2010; 7(6): 708-717; Sakuishi et al. JExp Med. 2010; 207(10): 2187-2194; Ngiow et al. Cancer Res. 2011;71(21): 6567-6571; Sakuishi et al Trends Immunol. 2011; 32(8): 345-349;Jing et al. J Immunother Cancer. 2015; 3(1):2; Asayama et al.Oncotarget. 2017; 8(51): 88904-88917). In fact, blockade of TIM-3 onmacrophages and antigen cross-presenting dendritic cells enhancesactivation and inflammatory cytokine/chemokine production (Zhang 2011;Zhang et al. (2012) J. Leukoc Biol 91(2):189-96; Chiba et al. (2012) NatImmunol. 13(9):832-42; de Mingo Pulido et al. (2018) Cancer Cell33(1):60-74), ultimately leading to enhanced effector T cells responses.

The combinations described herein include a TIM-3 inhibitor and can beused to treat a cancer, e.g., a hematological cancer. Combininghypomethylating agents with additional agents may improve their clinicalefficacy and overcome resistance. Preclinical data suggest thathypomethylating agents enhance checkpoint expression and that asynergistic response can be produced by using a checkpoint inhibitor anda hypomethylating agent. Hypomethylating agents induce increasedexpression of checkpoints molecules in MDS patients, e.g., TIM-3, PD-1,PD-L1, PD-L2 and CTLA4, potentially downregulating immune-mediatedanti-leukemic effects (Yang et al., (2014) Leukemia, 28(6):1280-8;Ørskov et al. (2015) Oncotarget, 6(11): 9612-9626). Additionally,demethylation of the TIM-3 promoter has been shown to be important forthe stable expression of TIM-3 on T-cells, indicating that modulation ofthe expression of TIM-3 by hypomethylating agents (e.g., azacitidine ordecitabine) can have important immunomodulatory implications (Chou etal. (2016) Genes Immun 17(3): 179-86). Without wishing to be bound bytheory, it is believed that in some embodiments, a combination describedherein (e.g., a combination comprising an anti-TIM-3 antibody moleculedescribed herein) can be used to decrease an immunosuppressive tumormicroenvironment.

Without wishing to be bound by theory, it is believed that in someembodiments, a combination comprising a TIM-3 inhibitor and ahypomethylating agent, can be administered safely, and that the TIM-3inhibitor can improve the efficacy of the hypomethylating agent, and/orimprove durability of response.

Accordingly, disclosed herein, at least in part, are combinationtherapies that can be used to treat or prevent disorders, such ascancerous disorders. In certain embodiments, the combination comprises aTIM-3 inhibitor and a hypomethylating agent. In some embodiments, theTIM-3 inhibitor comprises an antibody molecule (e.g., humanized antibodymolecule) that binds to TIM-3 with high affinity and specificity. Thecombinations described herein can be used according to a dosage regimendescribed herein. Pharmaceutical compositions and dose formulationsrelating to the combinations described herein are also provided.

Definitions

Additional terms are defined below and throughout the application.

As used herein, the articles “a” and “an” refer to one or to more thanone (e.g., to at least one) of the grammatical object of the article.

The term “or” is used herein to mean, and is used interchangeably with,the term “and/or,” unless context clearly indicates otherwise.

“About” and “approximately” shall generally mean an acceptable degree oferror for the quantity measured given the nature or precision of themeasurements. Exemplary degrees of error are within 20 percent (%),typically, within 10%, and more typically, within 5% of a given value orrange of values.

By “a combination” or “in combination with,” it is not intended to implythat the therapy or the therapeutic agents must be administered at thesame time and/or formulated for delivery together, although thesemethods of delivery are within the scope described herein. Thetherapeutic agents in the combination can be administered concurrentlywith, prior to, or subsequent to, one or more other additional therapiesor therapeutic agents. The therapeutic agents or therapeutic protocolcan be administered in any order. In general, each agent will beadministered at a dose and/or on a time schedule determined for thatagent. In will further be appreciated that the additional therapeuticagent utilized in this combination may be administered together in asingle composition or administered separately in different compositions.In general, it is expected that additional therapeutic agents utilizedin combination be utilized at levels that do not exceed the levels atwhich they are utilized individually. In some embodiments, the levelsutilized in combination will be lower than those utilized individually.

In embodiments, the additional therapeutic agent is administered at atherapeutic or lower-than therapeutic dose. In certain embodiments, theconcentration of the second therapeutic agent that is required toachieve inhibition, e.g., growth inhibition, is lower when the secondtherapeutic agent is administered in combination with the firsttherapeutic agent, e.g., the anti-TIM-3 antibody molecule, than when thesecond therapeutic agent is administered individually. In certainembodiments, the concentration of the first therapeutic agent that isrequired to achieve inhibition, e.g., growth inhibition, is lower whenthe first therapeutic agent is administered in combination with thesecond therapeutic agent than when the first therapeutic agent isadministered individually. In certain embodiments, in a combinationtherapy, the concentration of the second therapeutic agent that isrequired to achieve inhibition, e.g., growth inhibition, is lower thanthe therapeutic dose of the second therapeutic agent as a monotherapy,e.g., 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90%lower. In certain embodiments, in a combination therapy, theconcentration of the first therapeutic agent that is required to achieveinhibition, e.g., growth inhibition, is lower than the therapeutic doseof the first therapeutic agent as a monotherapy, e.g., 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, or 80-90% lower.

The term “inhibition,” “inhibitor,” or “antagonist” includes a reductionin a certain parameter, e.g., an activity, of a given molecule, e.g., animmune checkpoint inhibitor. For example, inhibition of an activity,e.g., a TIM-3 activity, of at least 5%, 10%, 20%, 30%, 40% or more isincluded by this term. Thus, inhibition need not be 100%.

The term “activation,” “activator,” or “agonist” includes an increase ina certain parameter, e.g., an activity, of a given molecule, e.g., acostimulatory molecule. For example, increase of an activity, e.g., acostimulatory activity, of at least 5%, 10%, 25%, 50%, 75% or more isincluded by this term.

The term “anti-cancer effect” refers to a biological effect which can bemanifested by various means, including but not limited to, e.g., adecrease in tumor volume, a decrease in the number of cancer cells, adecrease in the number of metastases, an increase in life expectancy,decrease in cancer cell proliferation, decrease in cancer cell survival,or amelioration of various physiological symptoms associated with thecancerous condition. An “anti-cancer effect” can also be manifested bythe ability of the peptides, polynucleotides, cells and antibodies inprevention of the occurrence of cancer in the first place.

The term “anti-tumor effect” refers to a biological effect which can bemanifested by various means, including but not limited to, e.g., adecrease in tumor volume, a decrease in the number of tumor cells, adecrease in tumor cell proliferation, or a decrease in tumor cellsurvival.

The term “cancer” refers to a disease characterized by the rapid anduncontrolled growth of aberrant cells. Cancer cells can spread locallyor through the bloodstream and lymphatic system to other parts of thebody. Examples of various cancers are described herein and include butare not limited to, solid tumors, e.g., lung cancer, breast cancer,prostate cancer, ovarian cancer, cervical cancer, skin cancer,pancreatic cancer, colorectal cancer, renal cancer, liver cancer, andbrain cancer, and hematologic malignancies, e.g., lymphoma and leukemia,and the like. The terms “tumor” and “cancer” are used interchangeablyherein, e.g., both terms encompass solid and liquid, e.g., diffuse orcirculating, tumors. As used herein, the term “cancer” or “tumor”includes premalignant, as well as malignant cancers and tumors.

The term “antigen presenting cell” or “APC” refers to an immune systemcell such as an accessory cell (e.g., a B-cell, a dendritic cell, andthe like) that displays a foreign antigen complexed with majorhistocompatibility complexes (MHC's) on its surface. T-cells mayrecognize these complexes using their T-cell receptors (TCRs). APCsprocess antigens and present them to T-cells.

The term “costimulatory molecule” refers to the cognate binding partneron a T cell that specifically binds with a costimulatory ligand, therebymediating a costimulatory response by the T cell, such as, but notlimited to, proliferation. Costimulatory molecules are cell surfacemolecules other than antigen receptors or their ligands that arerequired for an efficient immune response. Costimulatory moleculesinclude, but are not limited to, an MHC class I molecule, TNF receptorproteins, Immunoglobulin-like proteins, cytokine receptors, integrins,signalling lymphocytic activation molecules (SLAM proteins), activatingNK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27,CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a/CD18), 4-1BB (CD137), B7-H3,CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2,SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha,CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4,IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL,CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18,LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE/RANKL, DNAM1 (CD226), SLAMF4(CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160(BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM(SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS,SLP-76, PAG/Cbp, CD19a, and a ligand that specifically binds with CD83.

“Immune effector cell,” or “effector cell” as that term is used herein,refers to a cell that is involved in an immune response, e.g., in thepromotion of an immune effector response. Examples of immune effectorcells include T cells, e.g., alpha/beta T cells and gamma/delta T cells,B cells, natural killer (NK) cells, natural killer T (NKT) cells, mastcells, and myeloid-derived phagocytes.

“Immune effector” or “effector” “function” or “response,” as that termis used herein, refers to function or response, e.g., of an immuneeffector cell, that enhances or promotes an immune attack of a targetcell. E.g., an immune effector function or response refers a property ofa T or NK cell that promotes killing or the inhibition of growth orproliferation, of a target cell. In the case of a T cell, primarystimulation and co-stimulation are examples of immune effector functionor response.

The term “effector function” refers to a specialized function of a cell.Effector function of a T cell, for example, may be cytolytic activity orhelper activity including the secretion of cytokines.

As used herein, the terms “treat,” “treatment” and “treating” refer tothe reduction or amelioration of the progression, severity and/orduration of a disorder, e.g., a proliferative disorder, or theamelioration of one or more symptoms (preferably, one or morediscernible symptoms) of the disorder resulting from the administrationof one or more therapies. In specific embodiments, the terms “treat,”“treatment” and “treating” refer to the amelioration of at least onemeasurable physical parameter of a proliferative disorder, such asgrowth of a tumor, not necessarily discernible by the patient. In otherembodiments the terms “treat,” “treatment” and “treating” refer to theinhibition of the progression of a proliferative disorder, eitherphysically by, e.g., stabilization of a discernible symptom,physiologically by, e.g., stabilization of a physical parameter, orboth. In other embodiments the terms “treat,” “treatment” and “treating”refer to the reduction or stabilization of tumor size or cancerous cellcount.

The compositions, formulations, and methods of the present inventionencompass polypeptides and nucleic acids having the sequences specified,or sequences substantially identical or similar thereto, e.g., sequencesat least 85%, 90%, 95% identical or higher to the sequence specified. Inthe context of an amino acid sequence, the term “substantiallyidentical” is used herein to refer to a first amino acid that contains asufficient or minimum number of amino acid residues that are i)identical to, or ii) conservative substitutions of aligned amino acidresidues in a second amino acid sequence such that the first and secondamino acid sequences can have a common structural domain and/or commonfunctional activity. For example, amino acid sequences that contain acommon structural domain having at least about 85%, 90%. 91%, 92%, 93%,94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., asequence provided herein.

In the context of nucleotide sequence, the term “substantiallyidentical” is used herein to refer to a first nucleic acid sequence thatcontains a sufficient or minimum number of nucleotides that areidentical to aligned nucleotides in a second nucleic acid sequence suchthat the first and second nucleotide sequences encode a polypeptidehaving common functional activity, or encode a common structuralpolypeptide domain or a common functional polypeptide activity. Forexample, nucleotide sequences having at least about 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence,e.g., a sequence provided herein.

The term “functional variant” refers to polypeptides that have asubstantially identical amino acid sequence to the naturally-occurringsequence, or are encoded by a substantially identical nucleotidesequence, and are capable of having one or more activities of thenaturally-occurring sequence.

Calculations of homology or sequence identity between sequences (theterms are used interchangeably herein) are performed as follows.

To determine the percent identity of two amino acid sequences, or of twonucleic acid sequences, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in one or both of a first and asecond amino acid or nucleic acid sequence for optimal alignment andnon-homologous sequences can be disregarded for comparison purposes). Ina preferred embodiment, the length of a reference sequence aligned forcomparison purposes is at least 30%, preferably at least 40%, morepreferably at least 50%, 60%, and even more preferably at least 70%,80%, 90%, 100% of the length of the reference sequence. The amino acidresidues or nucleotides at corresponding amino acid positions ornucleotide positions are then compared. When a position in the firstsequence is occupied by the same amino acid residue or nucleotide as thecorresponding position in the second sequence, then the molecules areidentical at that position (as used herein amino acid or nucleic acid“identity” is equivalent to amino acid or nucleic acid “homology”).

The percent identity between the two sequences is a function of thenumber of identical positions shared by the sequences, taking intoaccount the number of gaps, and the length of each gap, which need to beintroduced for optimal alignment of the two sequences.

The comparison of sequences and determination of percent identitybetween two sequences can be accomplished using a mathematicalalgorithm. In a preferred embodiment, the percent identity between twoamino acid sequences is determined using the Needleman and Wunsch((1970) J. Mol. Biol. 48:444-453) algorithm which has been incorporatedinto the GAP program in the GCG software package (available at gcg.com),using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.In yet another preferred embodiment, the percent identity between twonucleotide sequences is determined using the GAP program in the GCGsoftware package (available at gcg.com), using a NWSgapdna.CMP matrixand a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2,3, 4, 5, or 6. A particularly preferred set of parameters (and the onethat should be used unless otherwise specified) are a Blossum 62 scoringmatrix with a gap penalty of 12, a gap extend penalty of 4, and aframeshift gap penalty of 5.

The percent identity between two amino acid or nucleotide sequences canbe determined using the algorithm of E. Meyers and W. Miller ((1989)CABIOS, 4:11-17) which has been incorporated into the ALIGN program(version 2.0), using a PAM120 weight residue table, a gap length penaltyof 12 and a gap penalty of 4.

The nucleic acid and protein sequences described herein can be used as a“query sequence” to perform a search against public databases, forexample, to identify other family members or related sequences. Suchsearches can be performed using the NBLAST and XBLAST programs (version2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLASTnucleotide searches can be performed with the NBLAST program, score=100,wordlength=12 to obtain nucleotide sequences homologous to a nucleicacid molecules of the invention. BLAST protein searches can be performedwith the XBLAST program, score=50, wordlength=3 to obtain amino acidsequences homologous to protein molecules of the invention. To obtaingapped alignments for comparison purposes, Gapped BLAST can be utilizedas described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402.When utilizing BLAST and Gapped BLAST programs, the default parametersof the respective programs (e.g., XBLAST and NBLAST) can be used. Seencbi.nlm.nih.gov.

As used herein, the term “hybridizes under low stringency, mediumstringency, high stringency, or very high stringency conditions”describes conditions for hybridization and washing. Guidance forperforming hybridization reactions can be found in Current Protocols inMolecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which isincorporated by reference. Aqueous and nonaqueous methods are describedin that reference and either can be used. Specific hybridizationconditions referred to herein are as follows: 1) low stringencyhybridization conditions in 6× sodium chloride/sodium citrate (SSC) atabout 45° C., followed by two washes in 0.2×SSC, 0.1% SDS at least at50° C. (the temperature of the washes can be increased to 55° C. for lowstringency conditions); 2) medium stringency hybridization conditions in6×SSC at about 45° C., followed by one or more washes in 0.2×SSC, 0.1%SDS at 60° C.; 3) high stringency hybridization conditions in 6×SSC atabout 45° C., followed by one or more washes in 0.2×SSC, 0.1% SDS at 65°C.; and preferably 4) very high stringency hybridization conditions are0.5M sodium phosphate, 7% SDS at 65° C., followed by one or more washesat 0.2×SSC, 1% SDS at 65° C. Very high stringency conditions (4) are thepreferred conditions and the ones that should be used unless otherwisespecified.

It is understood that the molecules of the present invention may haveadditional conservative or non-essential amino acid substitutions, whichdo not have a substantial effect on their functions.

The term “amino acid” is intended to embrace all molecules, whethernatural or synthetic, which include both an amino functionality and anacid functionality and capable of being included in a polymer ofnaturally-occurring amino acids. Exemplary amino acids includenaturally-occurring amino acids; analogs, derivatives and congenersthereof; amino acid analogs having variant side chains; and allstereoisomers of any of any of the foregoing. As used herein the term“amino acid” includes both the D- or L-optical isomers andpeptidomimetics.

A “conservative amino acid substitution” is one in which the amino acidresidue is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art. These families include amino acids with basicside chains (e.g., lysine, arginine, histidine), acidic side chains(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine),nonpolar side chains (e.g., alanine, valine, leucine, isoleucine,proline, phenylalanine, methionine, tryptophan), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine).

The terms “polypeptide,” “peptide” and “protein” (if single chain) areused interchangeably herein to refer to polymers of amino acids of anylength. The polymer may be linear or branched, it may comprise modifiedamino acids, and it may be interrupted by non-amino acids. The termsalso encompass an amino acid polymer that has been modified; forexample, disulfide bond formation, glycosylation, lipidation,acetylation, phosphorylation, or any other manipulation, such asconjugation with a labeling component. The polypeptide can be isolatedfrom natural sources, can be a produced by recombinant techniques from aeukaryotic or prokaryotic host, or can be a product of syntheticprocedures.

The terms “nucleic acid,” “nucleic acid sequence,” “nucleotidesequence,” or “polynucleotide sequence,” and “polynucleotide” are usedinterchangeably. They refer to a polymeric form of nucleotides of anylength, either deoxyribonucleotides or ribonucleotides, or analogsthereof. The polynucleotide may be either single-stranded ordouble-stranded, and if single-stranded may be the coding strand ornon-coding (antisense) strand. A polynucleotide may comprise modifiednucleotides, such as methylated nucleotides and nucleotide analogs. Thesequence of nucleotides may be interrupted by non-nucleotide components.A polynucleotide may be further modified after polymerization, such asby conjugation with a labeling component. The nucleic acid may be arecombinant polynucleotide, or a polynucleotide of genomic, cDNA,semisynthetic, or synthetic origin which either does not occur in natureor is linked to another polynucleotide in a nonnatural arrangement.

The term “isolated,” as used herein, refers to material that is removedfrom its original or native environment (e.g., the natural environmentif it is naturally occurring). For example, a naturally-occurringpolynucleotide or polypeptide present in a living animal is notisolated, but the same polynucleotide or polypeptide, separated by humanintervention from some or all of the co-existing materials in thenatural system, is isolated. Such polynucleotides could be part of avector and/or such polynucleotides or polypeptides could be part of acomposition, and still be isolated in that such vector or composition isnot part of the environment in which it is found in nature.

Various aspects of the invention are described in further detail below.Additional definitions are set out throughout the specification.

TIM-3 Inhibitors

In certain embodiments, the combination described herein includes aTIM-3 inhibitor, e.g., an anti-TIM-3 antibody molecule. In someembodiments, the anti-TIM-3 antibody molecule binds to a mammalian,e.g., human, TIM-3. For example, the antibody molecule bindsspecifically to an epitope, e.g., linear or conformational epitope onTIM-3.

As used herein, the term “antibody molecule” refers to a protein, e.g.,an immunoglobulin chain or fragment thereof, comprising at least oneimmunoglobulin variable domain sequence. The term “antibody molecule”includes, for example, a monoclonal antibody (including a full-lengthantibody which has an immunoglobulin Fc region). In an embodiment, anantibody molecule comprises a full-length antibody, or a full-lengthimmunoglobulin chain. In an embodiment, an antibody molecule comprisesan antigen binding or functional fragment of a full-length antibody, ora full-length immunoglobulin chain. In an embodiment, an antibodymolecule is a multispecific antibody molecule, e.g., it comprises aplurality of immunoglobulin variable domain sequences, wherein a firstimmunoglobulin variable domain sequence of the plurality has bindingspecificity for a first epitope and a second immunoglobulin variabledomain sequence of the plurality has binding specificity for a secondepitope. In an embodiment, a multispecific antibody molecule is abispecific antibody molecule.

In an embodiment, an antibody molecule is a monospecific antibodymolecule and binds a single epitope. For example, a monospecificantibody molecule can have a plurality of immunoglobulin variable domainsequences, each of which binds the same epitope.

In an embodiment, an antibody molecule is a multispecific antibodymolecule, e.g., it comprises a plurality of immunoglobulin variabledomains sequences, wherein a first immunoglobulin variable domainsequence of the plurality has binding specificity for a first epitopeand a second immunoglobulin variable domain sequence of the pluralityhas binding specificity for a second epitope. In an embodiment, thefirst and second epitopes are on the same antigen, e.g., the sameprotein (or subunit of a multimeric protein). In an embodiment, thefirst and second epitopes overlap. In an embodiment, the first andsecond epitopes do not overlap. In an embodiment, the first and secondepitopes are on different antigens, e.g., the different proteins (ordifferent subunits of a multimeric protein). In an embodiment, amultispecific antibody molecule comprises a third, fourth or fifthimmunoglobulin variable domain. In an embodiment, a multispecificantibody molecule is a bispecific antibody molecule, a trispecificantibody molecule, or tetraspecific antibody molecule,

In an embodiment, a multispecific antibody molecule is a bispecificantibody molecule. A bispecific antibody has specificity for no morethan two antigens. A bispecific antibody molecule is characterized by afirst immunoglobulin variable domain sequence which has bindingspecificity for a first epitope and a second immunoglobulin variabledomain sequence that has binding specificity for a second epitope. In anembodiment, the first and second epitopes are on the same antigen, e.g.,the same protein (or subunit of a multimeric protein). In an embodiment,the first and second epitopes overlap. In an embodiment the first andsecond epitopes do not overlap. In an embodiment, the first and secondepitopes are on different antigens, e.g., the different proteins (ordifferent subunits of a multimeric protein). In an embodiment, abispecific antibody molecule comprises a heavy chain variable domainsequence and a light chain variable domain sequence which have bindingspecificity for a first epitope and a heavy chain variable domainsequence and a light chain variable domain sequence which have bindingspecificity for a second epitope. In an embodiment, a bispecificantibody molecule comprises a half antibody having binding specificityfor a first epitope and a half antibody having binding specificity for asecond epitope. In an embodiment, a bispecific antibody moleculecomprises a half antibody, or fragment thereof, having bindingspecificity for a first epitope and a half antibody, or fragmentthereof, having binding specificity for a second epitope. In anembodiment, a bispecific antibody molecule comprises a scFv, or fragmentthereof, have binding specificity for a first epitope and a scFv, orfragment thereof, have binding specificity for a second epitope. In anembodiment, the first epitope is located on TIM-3 and the second epitopeis located on a PD-1, LAG-3, CEACAM (e.g., CEACAM-1 and/or CEACAM-5),PD-L1, or PD-L2.

Protocols for generating multi-specific (e.g., bispecific ortrispecific) or heterodimeric antibody molecules are known in the art;including but not limited to, for example, the “knob in a hole” approachdescribed in, e.g., U.S. Pat. No. 5,731,168; the electrostatic steeringFc pairing as described in, e.g., WO 09/089004, WO 06/106905 and WO2010/129304; Strand Exchange Engineered Domains (SEED) heterodimerformation as described in, e.g., WO 07/110205; Fab arm exchange asdescribed in, e.g., WO 08/119353, WO 2011/131746, and WO 2013/060867;double antibody conjugate, e.g., by antibody cross-linking to generate abi-specific structure using a heterobifunctional reagent having anamine-reactive group and a sulfhydryl reactive group as described in,e.g., U.S. Pat. No. 4,433,059; bispecific antibody determinantsgenerated by recombining half antibodies (heavy-light chain pairs orFabs) from different antibodies through cycle of reduction and oxidationof disulfide bonds between the two heavy chains, as described in, e.g.,U.S. Pat. No. 4,444,878; trifunctional antibodies, e.g., three Fab′fragments cross-linked through sulfhydryl reactive groups, as describedin, e.g., U.S. Pat. No. 5,273,743; biosynthetic binding proteins, e.g.,pair of scFvs cross-linked through C-terminal tails preferably throughdisulfide or amine-reactive chemical cross-linking, as described in,e.g., U.S. Pat. No. 5,534,254; bifunctional antibodies, e.g., Fabfragments with different binding specificities dimerized through leucinezippers (e.g., c-fos and c-jun) that have replaced the constant domain,as described in, e.g., U.S. Pat. No. 5,582,996; bispecific andoligospecific mono- and oligovalent receptors, e.g., VH-CH1 regions oftwo antibodies (two Fab fragments) linked through a polypeptide spacerbetween the CH1 region of one antibody and the VH region of the otherantibody typically with associated light chains, as described in, e.g.,U.S. Pat. No. 5,591,828; bispecific DNA-antibody conjugates, e.g.,crosslinking of antibodies or Fab fragments through a double strandedpiece of DNA, as described in, e.g., U.S. Pat. No. 5,635,602; bispecificfusion proteins, e.g., an expression construct containing two scFvs witha hydrophilic helical peptide linker between them and a full constantregion, as described in, e.g., U.S. Pat. No. 5,637,481; multivalent andmultispecific binding proteins, e.g., dimer of polypeptides having firstdomain with binding region of Ig heavy chain variable region, and seconddomain with binding region of Ig light chain variable region, generallytermed diabodies (higher order structures are also disclosed creatingbispecific, trispecific, or tetraspecific molecules, as described in,e.g., U.S. Pat. No. 5,837,242; minibody constructs with linked VL and VHchains further connected with peptide spacers to an antibody hingeregion and CH3 region, which can be dimerized to formbispecific/multivalent molecules, as described in, e.g., U.S. Pat. No.5,837,821; VH and VL domains linked with a short peptide linker (e.g., 5or 10 amino acids) or no linker at all in either orientation, which canform dimers to form bispecific diabodies; trimers and tetramers, asdescribed in, e.g., U.S. Pat. No. 5,844,094; String of VH domains (or VLdomains in family members) connected by peptide linkages withcrosslinkable groups at the C-terminus further associated with VLdomains to form a series of FVs (or scFvs), as described in, e.g., U.S.Pat. No. 5,864,019; and single chain binding polypeptides with both a VHand a VL domain linked through a peptide linker are combined intomultivalent structures through non-covalent or chemical crosslinking toform, e.g., homobivalent, heterobivalent, trivalent, and tetravalentstructures using both scFV or diabody type format, as described in,e.g., U.S. Pat. No. 5,869,620. Additional exemplary multispecific andbispecific molecules and methods of making the same are found, forexample, in U.S. Pat. Nos. 5,910,573, 5,932,448, 5,959,083, 5,989,830,6,005,079, 6,239,259, 6,294,353, 6,333,396, 6,476,198, 6,511,663,6,670,453, 6,743,896, 6,809,185, 6,833,441, 7,129,330, 7,183,076,7,521,056, 7,527,787, 7,534,866, 7,612,181, US 2002/004587A1, US2002/076406A1, US 2002/103345A1, US 2003/207346A1, US 2003/211078A1, US2004/219643A1, US 2004/220388A1, US 2004/242847A1, US 2005/003403A1, US2005/004352A1, US 2005/069552A1, US 2005/079170A1, US 2005/100543A1, US2005/136049A1, US 2005/136051A1, US 2005/163782A1, US 2005/266425A1, US2006/083747A1, US 2006/120960A1, US 2006/204493A1, US 2006/263367A1, US2007/004909A1, US 2007/087381A1, US 2007/128150A1, US 2007/141049A1, US2007/154901A1, US 2007/274985A1, US 2008/050370A1, US 2008/069820A1, US2008/152645A1, US 2008/171855A1, US 2008/241884A1, US 2008/254512A1, US2008/260738A1, US 2009/130106A1, US 2009/148905A1, US 2009/155275A1, US2009/162359A1, US 2009/162360A1, US 2009/175851A1, US 2009/175867A1, US2009/232811A1, US 2009/234105A1, US 2009/263392A1, US 2009/274649A1, EP346087A2, WO 00/06605A2, WO 02/072635A2, WO 04/081051A1, WO 06/020258A2,WO 2007/044887A2, WO 2007/095338A2, WO 2007/137760A2, WO 2008/119353A1,WO 2009/021754A2, WO 2009/068630A1, WO 91/03493A1, WO 93/23537A1, WO94/09131A1, WO 94/12625A2, WO 95/09917A1, WO 96/37621A2, WO 99/64460A1.The contents of the above-referenced applications are incorporatedherein by reference in their entireties.

In other embodiments, the anti-TIM-3 antibody molecule (e.g., amonospecific, bispecific, or multispecific antibody molecule) iscovalently linked, e.g., fused, to another partner e.g., a protein e.g.,one, two or more cytokines, e.g., as a fusion molecule for example afusion protein. In other embodiments, the fusion molecule comprises oneor more proteins, e.g., one, two or more cytokines. In one embodiment,the cytokine is an interleukin (IL) chosen from one, two, three or moreof IL-1, IL-2, IL-12, IL-15 or IL-21. In one embodiment, a bispecificantibody molecule has a first binding specificity to a first target(e.g., to PD-1), a second binding specificity to a second target (e.g.,LAG-3 or TIM-3), and is optionally linked to an interleukin (e.g.,IL-12) domain e.g., full length IL-12 or a portion thereof.

A “fusion protein” and a “fusion polypeptide” refer to a polypeptidehaving at least two portions covalently linked together, where each ofthe portions is a polypeptide having a different property. The propertymay be a biological property, such as activity in vitro or in vivo. Theproperty can also be simple chemical or physical property, such asbinding to a target molecule, catalysis of a reaction, etc. The twoportions can be linked directly by a single peptide bond or through apeptide linker, but are in reading frame with each other.

In an embodiment, an antibody molecule comprises a diabody, and asingle-chain molecule, as well as an antigen-binding fragment of anantibody (e.g., Fab, F(ab′)₂, and Fv). For example, an antibody moleculecan include a heavy (H) chain variable domain sequence (abbreviatedherein as VH), and a light (L) chain variable domain sequence(abbreviated herein as VL). In an embodiment an antibody moleculecomprises or consists of a heavy chain and a light chain (referred toherein as a half antibody. In another example, an antibody moleculeincludes two heavy (H) chain variable domain sequences and two light (L)chain variable domain sequence, thereby forming two antigen bindingsites, such as Fab, Fab′, F(ab′)₂, Fc, Fd, Fd′, Fv, single chainantibodies (scFv for example), single variable domain antibodies,diabodies (Dab) (bivalent and bispecific), and chimeric (e.g.,humanized) antibodies, which may be produced by the modification ofwhole antibodies or those synthesized de novo using recombinant DNAtechnologies. These functional antibody fragments retain the ability toselectively bind with their respective antigen or receptor. Antibodiesand antibody fragments can be from any class of antibodies including,but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass(e.g., IgG1, IgG2, IgG3, and IgG4) of antibodies. The preparation ofantibody molecules can be monoclonal or polyclonal. An antibody moleculecan also be a human, humanized, CDR-grafted, or in vitro generatedantibody. The antibody can have a heavy chain constant region chosenfrom, e.g., IgG1, IgG2, IgG3, or IgG4. The antibody can also have alight chain chosen from, e.g., kappa or lambda. The term“immunoglobulin” (Ig) is used interchangeably with the term “antibody”herein.

Examples of antigen-binding fragments of an antibody molecule include:(i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CLand CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprisingtwo Fab fragments linked by a disulfide bridge at the hinge region;(iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fvfragment consisting of the VL and VH domains of a single arm of anantibody, (v) a diabody (dAb) fragment, which consists of a VH domain;(vi) a camelid or camelized variable domain; (vii) a single chain Fv(scFv), see e.g., Bird et al. (1988) Science 242:423-426; and Huston etal. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) a singledomain antibody. These antibody fragments are obtained usingconventional techniques known to those with skill in the art, and thefragments are screened for utility in the same manner as are intactantibodies.

The term “antibody” includes intact molecules as well as functionalfragments thereof. Constant regions of the antibodies can be altered,e.g., mutated, to modify the properties of the antibody (e.g., toincrease or decrease one or more of: Fc receptor binding, antibodyglycosylation, the number of cysteine residues, effector cell function,or complement function).

Antibody molecules can also be single domain antibodies. Single domainantibodies can include antibodies whose complementary determiningregions are part of a single domain polypeptide. Examples include, butare not limited to, heavy chain antibodies, antibodies naturally devoidof light chains, single domain antibodies derived from conventional4-chain antibodies, engineered antibodies and single domain scaffoldsother than those derived from antibodies. Single domain antibodies maybe any of the art, or any future single domain antibodies. Single domainantibodies may be derived from any species including, but not limited tomouse, human, camel, llama, fish, shark, goat, rabbit, and bovine.According to another aspect of the invention, a single domain antibodyis a naturally occurring single domain antibody known as heavy chainantibody devoid of light chains. Such single domain antibodies aredisclosed in WO 94/04678, for example. For clarity reasons, thisvariable domain derived from a heavy chain antibody naturally devoid oflight chain is known herein as a VHH or nanobody to distinguish it fromthe conventional VH of four chain immunoglobulins. Such a VHH moleculecan be derived from antibodies raised in Camelidae species, for examplein camel, llama, dromedary, alpaca and guanaco. Other species besidesCamelidae may produce heavy chain antibodies naturally devoid of lightchain; such VHHs are within the scope of the invention.

The VH and VL regions can be subdivided into regions ofhypervariability, termed “complementarity determining regions” (CDR),interspersed with regions that are more conserved, termed “frameworkregions” (FR or FW).

The extent of the framework region and CDRs has been precisely definedby a number of methods (see, Kabat, E. A., et al. (1991) Sequences ofProteins of Immunological Interest, Fifth Edition, U.S. Department ofHealth and Human Services, NIH Publication No. 91-3242; Chothia, C. etal. (1987) J. Mol. Biol. 196:901-917; and the AbM definition used byOxford Molecular's AbM antibody modeling software. See, generally, e.g.,Protein Sequence and Structure Analysis of Antibody Variable Domains.In: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R.,Springer-Verlag, Heidelberg).

The terms “complementarity determining region,” and “CDR,” as usedherein refer to the sequences of amino acids within antibody variableregions which confer antigen specificity and binding affinity. Ingeneral, there are three CDRs in each heavy chain variable region(HCDR1, HCDR2, and HCDR3) and three CDRs in each light chain variableregion (LCDR1, LCDR2, and LCDR3).

The precise amino acid sequence boundaries of a given CDR can bedetermined using any of a number of well-known schemes, including thosedescribed by Kabat et al. (1991), “Sequences of Proteins ofImmunological Interest,” 5th Ed. Public Health Service, NationalInstitutes of Health, Bethesda, Md. (“Kabat” numbering scheme),Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering scheme).As used herein, the CDRs defined according the “Chothia” number schemeare also sometimes referred to as “hypervariable loops.”

For example, under Kabat, the CDR amino acid residues in the heavy chainvariable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and95-102 (HCDR3); and the CDR amino acid residues in the light chainvariable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and89-97 (LCDR3). Under Chothia the CDR amino acids in the VH are numbered26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acidresidues in VL are numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96(LCDR3). By combining the CDR definitions of both Kabat and Chothia, theCDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50-56(LCDR2), and 89-97 (LCDR3) in human VL.

Generally, unless specifically indicated, the anti-TIM-3 antibodymolecules can include any combination of one or more Kabat CDRs and/orChothia hypervariable loops, e.g., described in Table 7. In oneembodiment, the following definitions are used for the anti-TIM-3antibody molecules described in Table 7: HCDR1 according to the combinedCDR definitions of both Kabat and Chothia, and HCCDRs 2-3 and LCCDRs 1-3according the CDR definition of Kabat. Under all definitions, each VHand VL typically includes three CDRs and four FRs, arranged fromamino-terminus to carboxy-terminus in the following order: FR1, CDR1,FR2, CDR2, FR3, CDR3, FR4.

As used herein, an “immunoglobulin variable domain sequence” refers toan amino acid sequence which can form the structure of an immunoglobulinvariable domain. For example, the sequence may include all or part ofthe amino acid sequence of a naturally-occurring variable domain. Forexample, the sequence may or may not include one, two, or more N- orC-terminal amino acids, or may include other alterations that arecompatible with formation of the protein structure.

The term “antigen-binding site” refers to the part of an antibodymolecule that comprises determinants that form an interface that bindsto the TIM-3polypeptide, or an epitope thereof. With respect to proteins(or protein mimetics), the antigen-binding site typically includes oneor more loops (of at least four amino acids or amino acid mimics) thatform an interface that binds to the TIM-3 polypeptide. Typically, theantigen-binding site of an antibody molecule includes at least one ortwo CDRs and/or hypervariable loops, or more typically at least three,four, five or six CDRs and/or hypervariable loops.

The terms “compete” or “cross-compete” are used interchangeably hereinto refer to the ability of an antibody molecule to interfere withbinding of an anti-TIM-3 antibody molecule, e.g., an anti-TIM-3 antibodymolecule provided herein, to a target, e.g., human TIM-3. Theinterference with binding can be direct or indirect (e.g., through anallosteric modulation of the antibody molecule or the target). Theextent to which an antibody molecule is able to interfere with thebinding of another antibody molecule to the target, and thereforewhether it can be said to compete, can be determined using a competitionbinding assay, for example, a FACS assay, an ELISA or BIACORE assay. Insome embodiments, a competition binding assay is a quantitativecompetition assay. In some embodiments, a first anti-TIM-3 antibodymolecule is said to compete for binding to the target with a secondanti-TIM-3 antibody molecule when the binding of the first antibodymolecule to the target is reduced by 10% or more, e.g., 20% or more, 30%or more, 40% or more, 50% or more, 55% or more, 60% or more, 65% ormore, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more,95% or more, 98% or more, 99% or more in a competition binding assay(e.g., a competition assay described herein).

The terms “monoclonal antibody” or “monoclonal antibody composition” asused herein refer to a preparation of antibody molecules of singlemolecular composition. A monoclonal antibody composition displays asingle binding specificity and affinity for a particular epitope. Amonoclonal antibody can be made by hybridoma technology or by methodsthat do not use hybridoma technology (e.g., recombinant methods).

An “effectively human” protein is a protein that does not evoke aneutralizing antibody response, e.g., the human anti-murine antibody(HAMA) response. HAMA can be problematic in a number of circumstances,e.g., if the antibody molecule is administered repeatedly, e.g., intreatment of a chronic or recurrent disease condition. A HAMA responsecan make repeated antibody administration potentially ineffectivebecause of an increased antibody clearance from the serum (see e.g.,Saleh et al., Cancer Immunol. Immunother. 32:180-190 (1990)) and alsobecause of potential allergic reactions (see e.g., LoBuglio et al.,Hybridoma, 5:5117-5123 (1986)).

The antibody molecule can be a polyclonal or a monoclonal antibody. Inother embodiments, the antibody can be recombinantly produced, e.g.,produced by phage display or by combinatorial methods.

Phage display and combinatorial methods for generating antibodies areknown in the art (as described in, e.g., Ladner et al. U.S. Pat. No.5,223,409; Kang et al. International Publication No. WO 92/18619; Doweret al. International Publication No. WO 91/17271; Winter et al.International Publication WO 92/20791; Markland et al. InternationalPublication No. WO 92/15679; Breitling et al. International PublicationWO 93/01288; McCafferty et al. International Publication No. WO92/01047; Garrard et al. International Publication No. WO 92/09690;Ladner et al. International Publication No. WO 90/02 809; Fuchs et al.(1991) Bio/Technology 9:1370-1 372; Hay et al. (1992) Hum AntibodyHybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffthset al. (1993) EMBO J 12:725-734; Hawkins et al. (1992) J Mol Biol226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al.(1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio/Technology9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; andBarbas et al. (1991) PNAS 88:7978-7982, the contents of all of which areincorporated by reference herein).

In one embodiment, the antibody is a fully human antibody (e.g., anantibody made in a mouse which has been genetically engineered toproduce an antibody from a human immunoglobulin sequence), or anon-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g.,monkey), camel antibody. Preferably, the non-human antibody is a rodent(mouse or rat antibody). Methods of producing rodent antibodies areknown in the art.

Human monoclonal antibodies can be generated using transgenic micecarrying the human immunoglobulin genes rather than the mouse system.Splenocytes from these transgenic mice immunized with the antigen ofinterest are used to produce hybridomas that secrete human mAbs withspecific affinities for epitopes from a human protein (see, e.g., Woodet al. International Application WO 91/00906, Kucherlapati et al. PCTpublication WO 91/10741; Lonberg et al. International Application WO92/03918; Kay et al. International Application 92/03917; Lonberg, N. etal. 1994 Nature 368:856-859; Green, L. L. et al. 1994 Nature Genet.7:13-21; Morrison, S. L. et al. 1994 Proc. Natl. Acad. Sci. USA81:6851-6855; Bruggeman et al. 1993 Year Immunol 7:33-40; Tuaillon etal. 1993 PNAS 90:3720-3724; Bruggeman et al. 1991 Eur J Immunol21:1323-1326).

An antibody can be one in which the variable region, or a portionthereof, e.g., the CDRs, are generated in a non-human organism, e.g., arat or mouse. Chimeric, CDR-grafted, and humanized antibodies are withinthe invention. Antibodies generated in a non-human organism, e.g., a rator mouse, and then modified, e.g., in the variable framework or constantregion, to decrease antigenicity in a human are within the invention.

Chimeric antibodies can be produced by recombinant DNA techniques knownin the art (see Robinson et al., International Patent PublicationPCT/US86/02269; Akira, et al., European Patent Application 184,187;Taniguchi, M., European Patent Application 171,496; Morrison et al.,European Patent Application 173,494; Neuberger et al., InternationalApplication WO 86/01533; Cabilly et al. U.S. Pat. No. 4,816,567; Cabillyet al., European Patent Application 125,023; Better et al. (1988 Science240:1041-1043); Liu et al. (1987) PNAS 84:3439-3443; Liu et al., 1987,J. Immunol. 139:3521-3526; Sun et al. (1987) PNAS 84:214-218; Nishimuraet al., 1987, Canc. Res. 47:999-1005; Wood et al. (1985) Nature314:446-449; and Shaw et al., 1988, J. Natl Cancer Inst. 80:1553-1559).

A humanized or CDR-grafted antibody will have at least one or two butgenerally all three recipient CDRs (of heavy and or light immunoglobulinchains) replaced with a donor CDR. The antibody may be replaced with atleast a portion of a non-human CDR or only some of the CDRs may bereplaced with non-human CDRs. It is only necessary to replace the numberof CDRs required for binding of the humanized antibody to PD-1.Preferably, the donor will be a rodent antibody, e.g., a rat or mouseantibody, and the recipient will be a human framework or a humanconsensus framework. Typically, the immunoglobulin providing the CDRs iscalled the “donor” and the immunoglobulin providing the framework iscalled the “acceptor.” In one embodiment, the donor immunoglobulin is anon-human (e.g., rodent). The acceptor framework is anaturally-occurring (e.g., a human) framework or a consensus framework,or a sequence about 85% or higher, preferably 90%, 95%, 99% or higheridentical thereto.

As used herein, the term “consensus sequence” refers to the sequenceformed from the most frequently occurring amino acids (or nucleotides)in a family of related sequences (see e.g., Winnaker, From Genes toClones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family ofproteins, each position in the consensus sequence is occupied by theamino acid occurring most frequently at that position in the family. Iftwo amino acids occur equally frequently, either can be included in theconsensus sequence. A “consensus framework” refers to the frameworkregion in the consensus immunoglobulin sequence.

An antibody can be humanized by methods known in the art (see e.g.,Morrison, S. L., 1985, Science 229:1202-1207, by Oi et al., 1986,BioTechniques 4:214, and by Queen et al. U.S. Pat. Nos. 5,585,089,5,693,761 and 5,693,762, the contents of all of which are herebyincorporated by reference).

Humanized or CDR-grafted antibodies can be produced by CDR-grafting orCDR substitution, wherein one, two, or all CDRs of an immunoglobulinchain can be replaced. See e.g., U.S. Pat. No. 5,225,539; Jones et al.1986 Nature 321:552-525; Verhoeyan et al. 1988 Science 239:1534; Beidleret al. 1988 J. Immunol. 141:4053-4060; Winter U.S. Pat. No. 5,225,539,the contents of all of which are hereby expressly incorporated byreference. Winter describes a CDR-grafting method which may be used toprepare the humanized antibodies of the present invention (UK PatentApplication GB 2188638A, filed on Mar. 26, 1987; Winter U.S. Pat. No.5,225,539), the contents of which is expressly incorporated byreference.

Also within the scope of the invention are humanized antibodies in whichspecific amino acids have been substituted, deleted or added. Criteriafor selecting amino acids from the donor are described in U.S. Pat. No.5,585,089, e.g., columns 12-16 of U.S. Pat. No. 5,585,089, e.g., columns12-16 of U.S. Pat. No. 5,585,089, the contents of which are herebyincorporated by reference. Other techniques for humanizing antibodiesare described in Padlan et al. EP 519596 A1, published on Dec. 23, 1992.

The antibody molecule can be a single chain antibody. A single-chainantibody (scFV) may be engineered (see, for example, Colcher, D. et al.(1999) Ann NY Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res2:245-52). The single chain antibody can be dimerized or multimerized togenerate multivalent antibodies having specificities for differentepitopes of the same target protein.

In yet other embodiments, the antibody molecule has a heavy chainconstant region chosen from, e.g., the heavy chain constant regions ofIgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; particularly,chosen from, e.g., the (e.g., human) heavy chain constant regions ofIgG1, IgG2, IgG3, and IgG4. In another embodiment, the antibody moleculehas a light chain constant region chosen from, e.g., the (e.g., human)light chain constant regions of kappa or lambda. The constant region canbe altered, e.g., mutated, to modify the properties of the antibody(e.g., to increase or decrease one or more of: Fc receptor binding,antibody glycosylation, the number of cysteine residues, effector cellfunction, and/or complement function). In one embodiment the antibodyhas: effector function; and can fix complement. In other embodiments theantibody does not; recruit effector cells; or fix complement. In anotherembodiment, the antibody has reduced or no ability to bind an Fcreceptor. For example, it is a isotype or subtype, fragment or othermutant, which does not support binding to an Fc receptor, e.g., it has amutagenized or deleted Fc receptor binding region.

Methods for altering an antibody constant region are known in the art.Antibodies with altered function, e.g. altered affinity for an effectorligand, such as FcR on a cell, or the C1 component of complement can beproduced by replacing at least one amino acid residue in the constantportion of the antibody with a different residue (see e.g., EP 388,151A1, U.S. Pat. Nos. 5,624,821 and 5,648,260, the contents of all of whichare hereby incorporated by reference) Similar type of alterations couldbe described which if applied to the murine, or other speciesimmunoglobulin would reduce or eliminate these functions.

An antibody molecule can be derivatized or linked to another functionalmolecule (e.g., another peptide or protein). As used herein, a“derivatized” antibody molecule is one that has been modified. Methodsof derivatization include but are not limited to the addition of afluorescent moiety, a radionucleotide, a toxin, an enzyme or an affinityligand such as biotin. Accordingly, the antibody molecules of theinvention are intended to include derivatized and otherwise modifiedforms of the antibodies described herein, including immunoadhesionmolecules. For example, an antibody molecule can be functionally linked(by chemical coupling, genetic fusion, noncovalent association orotherwise) to one or more other molecular entities, such as anotherantibody (e.g., a bispecific antibody or a diabody), a detectable agent,a cytotoxic agent, a pharmaceutical agent, and/or a protein or peptidethat can mediate association of the antibody or antibody portion withanother molecule (such as a streptavidin core region or a polyhistidinetag).

One type of derivatized antibody molecule is produced by crosslinkingtwo or more antibodies (of the same type or of different types, e.g., tocreate bispecific antibodies). Suitable crosslinkers include those thatare heterobifunctional, having two distinctly reactive groups separatedby an appropriate spacer (e.g., m-maleimidobenzoyl-N-hydroxysuccinimideester) or homobifunctional (e.g., disuccinimidyl suberate). Such linkersare available from Pierce Chemical Company, Rockford, Ill.

Useful detectable agents with which an antibody molecule of theinvention may be derivatized (or labeled) to include fluorescentcompounds, various enzymes, prosthetic groups, luminescent materials,bioluminescent materials, fluorescent emitting metal atoms, e.g.,europium (Eu), and other anthanides, and radioactive materials(described below). Exemplary fluorescent detectable agents includefluorescein, fluorescein isothiocyanate, rhodamine,5dimethylamine-1-napthalenesulfonyl chloride, phycoerythrin and thelike. An antibody may also be derivatized with detectable enzymes, suchas alkaline phosphatase, horseradish peroxidase, β-galactosidase,acetylcholinesterase, glucose oxidase and the like. When an antibody isderivatized with a detectable enzyme, it is detected by addingadditional reagents that the enzyme uses to produce a detectablereaction product. For example, when the detectable agent horseradishperoxidase is present, the addition of hydrogen peroxide anddiaminobenzidine leads to a colored reaction product, which isdetectable. An antibody molecule may also be derivatized with aprosthetic group (e.g., streptavidin/biotin and avidin/biotin). Forexample, an antibody may be derivatized with biotin, and detectedthrough indirect measurement of avidin or streptavidin binding. Examplesof suitable fluorescent materials include umbelliferone, fluorescein,fluorescein isothiocyanate, rhodamine, dichlorotriazinylaminefluorescein, dansyl chloride or phycoerythrin; an example of aluminescent material includes luminol; and examples of bioluminescentmaterials include luciferase, luciferin, and aequorin.

Labeled antibody molecule can be used, for example, diagnosticallyand/or experimentally in a number of contexts, including (i) to isolatea predetermined antigen by standard techniques, such as affinitychromatography or immunoprecipitation; (ii) to detect a predeterminedantigen (e.g., in a cellular lysate or cell supernatant) in order toevaluate the abundance and pattern of expression of the protein; (iii)to monitor protein levels in tissue as part of a clinical testingprocedure, e.g., to determine the efficacy of a given treatment regimen.

An antibody molecule may be conjugated to another molecular entity,typically a label or a therapeutic (e.g., a cytotoxic or cytostatic)agent or moiety. Radioactive isotopes can be used in diagnostic ortherapeutic applications.

The invention provides radiolabeled antibody molecules and methods oflabeling the same. In one embodiment, a method of labeling an antibodymolecule is disclosed. The method includes contacting an antibodymolecule, with a chelating agent, to thereby produce a conjugatedantibody.

As is discussed above, the antibody molecule can be conjugated to atherapeutic agent. Therapeutically active radioisotopes have alreadybeen mentioned. Examples of other therapeutic agents include taxol,cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin,etoposide, tenoposide, vincristine, vinblastine, colchicine,doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone,mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids,procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids,e.g., maytansinol (see, e.g., U.S. Pat. No. 5,208,020), CC-1065 (see,e.g., U.S. Pat. Nos. 5,475,092, 5,585,499, 5,846, 545) and analogs orhomologs thereof. Therapeutic agents include, but are not limited to,antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine,cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g.,mechlorethamine, thioepa chlorambucil, CC-1065, melphalan, carmustine(BSNU) and lomustine (CCNU), cyclothosphamide, busulfan,dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamineplatinum (II) (DDP) cisplatin), anthracyclinies (e.g., daunorubicin(formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin(formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)),and anti-mitotic agents (e.g., vincristine, vinblastine, taxol andmaytansinoids).

In one aspect, the disclosure provides a method of providing a targetbinding molecule that specifically binds to a target disclosed herein,e.g., TIM-3. For example, the target binding molecule is an antibodymolecule. The method includes: providing a target protein that comprisesat least a portion of non-human protein, the portion being homologous to(at least 70, 75, 80, 85, 87, 90, 92, 94, 95, 96, 97, 98% identical to)a corresponding portion of a human target protein, but differing by atleast one amino acid (e.g., at least one, two, three, four, five, six,seven, eight, or nine amino acids); obtaining an antibody molecule thatspecifically binds to the antigen; and evaluating efficacy of thebinding agent in modulating activity of the target protein. The methodcan further include administering the binding agent (e.g., antibodymolecule) or a derivative (e.g., a humanized antibody molecule) to ahuman subject.

This disclosure provides an isolated nucleic acid molecule encoding theabove antibody molecule, vectors and host cells thereof. The nucleicacid molecule includes but is not limited to RNA, genomic DNA and cDNA.

Exemplary TIM-3 Inhibitors

In certain embodiments, the combination described herein comprises ananti-TIM-3 antibody molecule. In one embodiment, the anti-TIM-3 antibodymolecule is disclosed in US 2015/0218274, published on Aug. 6, 2015,entitled “Antibody Molecules to TIM-3 and Uses Thereof,” incorporated byreference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule comprises at leastone, two, three, four, five or six complementarity determining regions(CDRs) (or collectively all of the CDRs) from a heavy and light chainvariable region comprising an amino acid sequence shown in Table 7(e.g., from the heavy and light chain variable region sequences ofABTIM3-hum11 or ABTIM3-hum03 disclosed in Table 7), or encoded by anucleotide sequence shown in Table 7. In some embodiments, the CDRs areaccording to the Kabat definition (e.g., as set out in Table 7). In someembodiments, the CDRs are according to the Chothia definition (e.g., asset out in Table 7). In one embodiment, one or more of the CDRs (orcollectively all of the CDRs) have one, two, three, four, five, six ormore changes, e.g., amino acid substitutions (e.g., conservative aminoacid substitutions) or deletions, relative to an amino acid sequenceshown in Table 7, or encoded by a nucleotide sequence shown in Table 7.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavychain variable region (VH) comprising a VHCDR1 amino acid sequence ofSEQ ID NO: 801, a VHCDR2 amino acid sequence of SEQ ID NO: 802, and aVHCDR3 amino acid sequence of SEQ ID NO: 803; and a light chain variableregion (VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 810, aVLCDR2 amino acid sequence of SEQ ID NO: 811, and a VLCDR3 amino acidsequence of SEQ ID NO: 812, each disclosed in Table 7. In oneembodiment, the anti-TIM-3 antibody molecule comprises a heavy chainvariable region (VH) comprising a VHCDR1 amino acid sequence of SEQ IDNO: 801, a VHCDR2 amino acid sequence of SEQ ID NO: 820, and a VHCDR3amino acid sequence of SEQ ID NO: 803; and a light chain variable region(VL) comprising a VLCDR1 amino acid sequence of SEQ ID NO: 810, a VLCDR2amino acid sequence of SEQ ID NO: 811, and a VLCDR3 amino acid sequenceof SEQ ID NO: 812, each disclosed in Table 7.

In one embodiment, the anti-TIM-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 806, or an amino acidsequence at least 85%, 90%, 95%, or 99% identical or higher to SEQ IDNO: 806. In one embodiment, the anti-TIM-3 antibody molecule comprises aVL comprising the amino acid sequence of SEQ ID NO: 816, or an aminoacid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQID NO: 816. In one embodiment, the anti-TIM-3 antibody moleculecomprises a VH comprising the amino acid sequence of SEQ ID NO: 822, oran amino acid sequence at least 85%, 90%, 95%, or 99% identical orhigher to SEQ ID NO: 822. In one embodiment, the anti-TIM-3 antibodymolecule comprises a VL comprising the amino acid sequence of SEQ ID NO:826, or an amino acid sequence at least 85%, 90%, 95%, or 99% identicalor higher to SEQ ID NO: 826. In one embodiment, the anti-TIM-3 antibodymolecule comprises a VH comprising the amino acid sequence of SEQ ID NO:806 and a VL comprising the amino acid sequence of SEQ ID NO: 816. Inone embodiment, the anti-TIM-3 antibody molecule comprises a VHcomprising the amino acid sequence of SEQ ID NO: 822 and a VL comprisingthe amino acid sequence of SEQ ID NO: 826.

In one embodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 807, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 807. In oneembodiment, the antibody molecule comprises a VL encoded by thenucleotide sequence of SEQ ID NO: 817, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 817. In oneembodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 823, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 823. In oneembodiment, the antibody molecule comprises a VL encoded by thenucleotide sequence of SEQ ID NO: 827, or a nucleotide sequence at least85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 827. In oneembodiment, the antibody molecule comprises a VH encoded by thenucleotide sequence of SEQ ID NO: 807 and a VL encoded by the nucleotidesequence of SEQ ID NO: 817. In one embodiment, the antibody moleculecomprises a VH encoded by the nucleotide sequence of SEQ ID NO: 823 anda VL encoded by the nucleotide sequence of SEQ ID NO: 827.

In one embodiment, the anti-TIM-3 antibody molecule comprises a heavychain comprising the amino acid sequence of SEQ ID NO: 808, or an aminoacid sequence at least 85%, 90%, 95%, or 99% identical or higher to SEQID NO: 808. In one embodiment, the anti-TIM-3 antibody moleculecomprises a light chain comprising the amino acid sequence of SEQ ID NO:818, or an amino acid sequence at least 85%, 90%, 95%, or 99% identicalor higher to SEQ ID NO: 818. In one embodiment, the anti-TIM-3 antibodymolecule comprises a heavy chain comprising the amino acid sequence ofSEQ ID NO: 824, or an amino acid sequence at least 85%, 90%, 95%, or 99%identical or higher to SEQ ID NO: 824. In one embodiment, the anti-TIM-3antibody molecule comprises a light chain comprising the amino acidsequence of SEQ ID NO: 828, or an amino acid sequence at least 85%, 90%,95%, or 99% identical or higher to SEQ ID NO: 828. In one embodiment,the anti-TIM-3 antibody molecule comprises a heavy chain comprising theamino acid sequence of SEQ ID NO: 808 and a light chain comprising theamino acid sequence of SEQ ID NO: 818. In one embodiment, the anti-TIM-3antibody molecule comprises a heavy chain comprising the amino acidsequence of SEQ ID NO: 824 and a light chain comprising the amino acidsequence of SEQ ID NO: 828.

In one embodiment, the antibody molecule comprises a heavy chain encodedby the nucleotide sequence of SEQ ID NO: 809, or a nucleotide sequenceat least 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 809. Inone embodiment, the antibody molecule comprises a light chain encoded bythe nucleotide sequence of SEQ ID NO: 819, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 819. Inone embodiment, the antibody molecule comprises a heavy chain encoded bythe nucleotide sequence of SEQ ID NO: 825, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 825. Inone embodiment, the antibody molecule comprises a light chain encoded bythe nucleotide sequence of SEQ ID NO: 829, or a nucleotide sequence atleast 85%, 90%, 95%, or 99% identical or higher to SEQ ID NO: 829. Inone embodiment, the antibody molecule comprises a heavy chain encoded bythe nucleotide sequence of SEQ ID NO: 809 and a light chain encoded bythe nucleotide sequence of SEQ ID NO: 819. In one embodiment, theantibody molecule comprises a heavy chain encoded by the nucleotidesequence of SEQ ID NO: 825 and a light chain encoded by the nucleotidesequence of SEQ ID NO: 829.

The antibody molecules described herein can be made by vectors, hostcells, and methods described in US 2015/0218274, incorporated byreference in its entirety.

TABLE 7Amino acid and nucleotide sequences of exemplary anti-TIM-3 antibody moleculesABTIM3-hum11 SEQ ID NO: 801 (Kabat) HCDR1 SYNMH SEQ ID NO:802 (Kabat)HCDR2 DIYPGNGDTSYNQKFKG SEQ ID NO: 803 (Kabat) HCDR3 VGGAFPMDYSEQ ID NO: 804 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 805 (Chothia) HCDR2YPGNGD SEQ ID NO: 803 (Chothia) HCDR3 VGGAFPMDY SEQ ID NO: 806 VHQVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSS SEQ ID NO: 807 DNA VHCAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTTTCTTGTAAAGCTAGTGGCTACACCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCAC TACCGTGACCGTGTCTAGCSEQ ID NO: 808 Heavy QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYNMHWVRQAPG chainQGLEWMGDIYPGNGDTSYNQKFKGRVTITADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLG SEQ ID NO: 809 DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACC heavyCGGCTCTAGCGTGAAAGTTTCTTGTAAAGCTAGTGGCTACAC chainCTTCACTAGCTATAATATGCACTGGGTTCGCCAGGCCCCAGGGCAAGGCCTCGAGTGGATGGGCGATATCTACCCCGGGAACGGCGACACTAGTTATAATCAGAAGTTTAAGGGTAGAGTCACTATCACCGCCGATAAGTCTACTAGCACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCTATGGACTACTGGGGTCAAGGCACTACCGTGACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACACTCAGAAGTCCCTGTCCCTCTCCCTGGGA SEQ ID NO: 810 (Kabat) LCDR1RASESVEYYGTSLMQ SEQ ID NO: 811 (Kabat) LCDR2 AASNVESSEQ ID NO: 812 (Kabat) LCDR3 QQSRKDPST SEQ ID NO: 813 (Chothia) LCDR1SESVEYYGTSL SEQ ID NO: 814 (Chothia) LCDR2 AAS SEQ ID NO: 815 (Chothia)LCDR3 SRKDPS SEQ ID NO: 816 VLAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKPGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATY FCQQSRKDPSTFGGGTKVEIKSEQ ID NO: 817 DNA VL GCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 818 LightAIQLTQSPSSLSASVGDRVTITCRASESVEYYGTSLMQWYQQKP chainGKAPKLLIYAASNVESGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 819 DNA lightGCTATTCAGCTGACTCAGTCACCTAGTAGCCTGAGCGCTAGT chainGTGGGCGATAGAGTGACTATCACCTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGGAAAGCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTTCTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGA CCAAGAGCTTCAACAGGGGCGAGTGCABTIM3-hum03 SEQ ID NO: 801 (Kabat) HCDR1 SYNMH SEQ ID NO: 820 (Kabat)HCDR2 DIYPGQGDTSYNQKFKG SEQ ID NO: 803 (Kabat) HCDR3 VGGAFPMDYSEQ ID NO: 804 (Chothia) HCDR1 GYTFTSY SEQ ID NO: 821 (Chothia) HCDR2YPGQGD SEQ ID NO: 803 (Chothia) HCDR3 VGGAFPMDY SEQ ID NO: 822 VHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSS SEQ ID NO: 823 DNA VHCAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCGCTAGTGTGAAAGTTAGCTGTAAAGCTAGTGGCTATACTTTCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCC TGGTCACCGTGTCTAGCSEQ ID NO: 824 Heavy QVQLVQSGAEVKKPGASVKVSCKASGYFTSYNMHWVRQAPG chainQGLEWIGDIYPGQGDTSYNQKFKGRATMTADKSTSTVYMELSSLRSEDTAVYYCARVGGAFPMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQ KSLSLSLG SEQ ID NO: 825 DNACAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACC heavyCGGCGCTAGTGTGAAAGTTAGCTGTAAAGCTAGTGGCTATAC chainTTTCACTTCTTATAATATGCACTGGGTCCGCCAGGCCCCAGGTCAAGGCCTCGAGTGGATCGGCGATATCTACCCCGGTCAAGGCGACACTTCCTATAATCAGAAGTTTAAGGGTAGAGCTACTATGACCGCCGATAAGTCTACTTCTACCGTCTATATGGAACTGAGTTCCCTGAGGTCTGAGGACACCGCCGTCTACTACTGCGCTAGAGTGGGCGGAGCCTTCCCAATGGACTACTGGGGTCAAGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCGTCCGTGTTCCCCCTGGCACCTTGTAGCCGGAGCACTAGCGAATCCACCGCTGCCCTCGGCTGCCTGGTCAAGGATTACTTCCCGGAGCCCGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCTCCGGAGTGCACACCTTCCCCGCTGTGCTGCAGAGCTCCGGGCTGTACTCGCTGTCGTCGGTGGTCACGGTGCCTTCATCTAGCCTGGGTACCAAGACCTACACTTGCAACGTGGACCACAAGCCTTCCAACACTAAGGTGGACAAGCGCGTCGAATCGAAGTACGGCCCACCGTGCCCGCCTTGTCCCGCGCCGGAGTTCCTCGGCGGTCCCTCGGTCTTTCTGTTCCCACCGAAGCCCAAGGACACTTTGATGATTTCCCGCACCCCTGAAGTGACATGCGTGGTCGTGGACGTGTCACAGGAAGATCCGGAGGTGCAGTTCAATTGGTACGTGGATGGCGTCGAGGTGCACAACGCCAAAACCAAGCCGAGGGAGGAGCAGTTCAACTCCACTTACCGCGTCGTGTCCGTGCTGACGGTGCTGCATCAGGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGGACTTCCTAGCTCAATCGAAAAGACCATCTCGAAAGCCAAGGGACAGCCCCGGGAACCCCAAGTGTATACCCTGCCACCGAGCCAGGAAGAAATGACTAAGAACCAAGTCTCATTGACTTGCCTTGTGAAGGGCTTCTACCCATCGGATATCGCCGTGGAATGGGAGTCCAACGGCCAGCCGGAAAACAACTACAAGACCACCCCTCCGGTGCTGGACTCAGACGGATCCTTCTTCCTCTACTCGCGGCTGACCGTGGATAAGAGCAGATGGCAGGAGGGAAATGTGTTCAGCTGTTCTGTGATGCATGAAGCCCTGCACAACCACTACAC TCAGAAGTCCCTGTCCCTCTCCCTGGGASEQ ID NO: 810 (Kabat) LCDR1 RASESVEYYGTSLMQ SEQ ID NO: 811 (Kabat)LCDR2 AASNVES SEQ ID NO: 812 (Kabat) LCDR3 QQSRKDPSTSEQ ID NO: 813 (Chothia) LCDR1 SESVEYYGTSL SEQ ID NO: 814 (Chothia)LCDR2 AAS SEQ ID NO: 815 (Chothia) LCDR3 SRKDPS SEQ ID NO: 826 VLDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKPGQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAV YYCQQSRKDPSTFGGGTKVEIKSEQ ID NO: 827 DNA VL GATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGCCTGGGCGAGCGGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAG SEQ ID NO: 828 LightDIVLTQSPDSLAVSLGERATINCRASESVEYYGTSLMQWYQQKP chainGQPPKLLIYAASNVESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQSRKDPSTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 829 DNA lightGATATCGTCCTGACTCAGTCACCCGATAGCCTGGCCGTCAGC chainCTGGGCGAGCGGGCTACTATTAACTGTAGAGCTAGTGAATCAGTCGAGTACTACGGCACTAGCCTGATGCAGTGGTATCAGCAGAAGCCCGGTCAACCCCCTAAGCTGCTGATCTACGCCGCCTCTAACGTGGAATCAGGCGTGCCCGATAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATTAGTAGCCTGCAGGCCGAGGACGTGGCCGTCTACTACTGTCAGCAGTCTAGGAAGGACCCTAGCACCTTCGGCGGAGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGA CCAAGAGCTTCAACAGGGGCGAGTGC

In one embodiment, the anti-TIM-3 antibody molecule includes at leastone or two heavy chain variable domain (optionally including a constantregion), at least one or two light chain variable domain (optionallyincluding a constant region), or both, comprising the amino acidsequence of ABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03,ABTIM3-hum04, ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08,ABTIM3-hum09, ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13,ABTIM3-hum14, ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18,ABTIM3-hum19, ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; oras described in Tables 1-4 of US 2015/0218274; or encoded by thenucleotide sequence in Tables 1-4; or a sequence substantially identical(e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higheridentical) to any of the aforesaid sequences. The anti-TIM-3 antibodymolecule, optionally, comprises a leader sequence from a heavy chain, alight chain, or both, as shown in US 2015/0218274; or a sequencesubstantially identical thereto.

In yet another embodiment, the anti-TIM-3 antibody molecule includes atleast one, two, or three complementarity determining regions (CDRs) froma heavy chain variable region and/or a light chain variable region of anantibody described herein, e.g., an antibody chosen from any of ABTIM3,ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04, ABTIM3-hum05,ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09, ABTIM3-hum10,ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14, ABTIM3-hum15,ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19, ABTIM3-hum20,ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23; or as described in Tables 1-4of US 2015/0218274; or encoded by the nucleotide sequence in Tables 1-4;or a sequence substantially identical (e.g., at least 80%, 85%, 90%,92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaidsequences.

In yet another embodiment, the anti-TIM-3 antibody molecule includes atleast one, two, or three CDRs (or collectively all of the CDRs) from aheavy chain variable region comprising an amino acid sequence shown inTables 1-4 of US 2015/0218274, or encoded by a nucleotide sequence shownin Tables 1-4. In one embodiment, one or more of the CDRs (orcollectively all of the CDRs) have one, two, three, four, five, six ormore changes, e.g., amino acid substitutions or deletions, relative tothe amino acid sequence shown in Tables 1-4, or encoded by a nucleotidesequence shown in Table 1-4.

In yet another embodiment, the anti-TIM-3 antibody molecule includes atleast one, two, or three CDRs (or collectively all of the CDRs) from alight chain variable region comprising an amino acid sequence shown inTables 1-4 of US 2015/0218274, or encoded by a nucleotide sequence shownin Tables 1-4. In one embodiment, one or more of the CDRs (orcollectively all of the CDRs) have one, two, three, four, five, six ormore changes, e.g., amino acid substitutions or deletions, relative tothe amino acid sequence shown in Tables 1-4, or encoded by a nucleotidesequence shown in Tables 1-4. In certain embodiments, the anti-TIM-3antibody molecule includes a substitution in a light chain CDR, e.g.,one or more substitutions in a CDR1, CDR2 and/or CDR3 of the lightchain.

In another embodiment, the anti-TIM-3 antibody molecule includes atleast one, two, three, four, five or six CDRs (or collectively all ofthe CDRs) from a heavy and light chain variable region comprising anamino acid sequence shown in Tables 1-4 of US 2015/0218274, or encodedby a nucleotide sequence shown in Tables 1-4. In one embodiment, one ormore of the CDRs (or collectively all of the CDRs) have one, two, three,four, five, six or more changes, e.g., amino acid substitutions ordeletions, relative to the amino acid sequence shown in Tables 1-4, orencoded by a nucleotide sequence shown in Tables 1-4.

In another embodiment, the anti-TIM-3 antibody molecule is MBG453.Without wising to be bound by theory, it is typically believed thatMBG453 is a high-affinity, ligand-blocking, humanized anti-TIM-3 IgG4antibody which can block the binding of TIM-3 to phosphatidyserine(PtdSer).

Other Exemplary TIM-3 Inhibitors

In one embodiment, the anti-TIM-3 antibody molecule is TSR-022(AnaptysBio/Tesaro). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of TSR-022. In oneembodiment, the anti-TIM-3 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain or light chain variable region sequence, or the heavy chain orlight chain sequence of APE5137 or APE5121, e.g., as disclosed in Table8. APE5137, APE5121, and other anti-TIM-3 antibodies are disclosed in WO2016/161270, incorporated by reference in its entirety.

In one embodiment, the anti-TIM-3 antibody molecule is the antibodyclone F38-2E2. In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of F38-2E2.

In one embodiment, the anti-TIM-3 antibody molecule is LY3321367 (EliLilly). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of LY3321367.

In one embodiment, the anti-TIM-3 antibody molecule is Sym023(Symphogen). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of Sym023.

In one embodiment, the anti-TIM-3 antibody molecule is BGB-A425(Beigene). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of BGB-A425.

In one embodiment, the anti-TIM-3 antibody molecule is INCAGN-2390(Agenus/Incyte). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of INCAGN-2390.

In one embodiment, the anti-TIM-3 antibody molecule is MBS-986258(BMS/Five Prime). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of MBS-986258.

In one embodiment, the anti-TIM-3 antibody molecule is RO-7121661(Roche). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of RO-7121661.

In one embodiment, the anti-TIM-3 antibody molecule is LY-3415244 (EliLilly). In one embodiment, the anti-TIM-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain variable region sequence and/or light chainvariable region sequence, or the heavy chain sequence and/or light chainsequence of LY-3415244.

In one embodiment, the anti-TIM-3 antibody molecule is BC-3402 (WuxiZhikanghongyi Biotechnology). In one embodiment, the anti-TIM-3 antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain variable region sequence and/orlight chain variable region sequence, or the heavy chain sequence and/orlight chain sequence of BC-3402.

In one embodiment, the anti-TIM-3 antibody molecule is SHR-1702(Medicine Co Ltd.). In one embodiment, the anti-TIM-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain variable region sequence and/or lightchain variable region sequence, or the heavy chain sequence and/or lightchain sequence of SHR-1702. SHR-1702 is disclosed, e.g., in WO2020/038355.

Further known anti-TIM-3 antibodies include those described, e.g., in WO2016/111947, WO 2016/071448, WO 2016/144803, U.S. Pat. Nos. 8,552,156,8,841,418, and 9,163,087, incorporated by reference in their entirety.

In one embodiment, the anti-TIM-3 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on TIM-3 as, one ofthe anti-TIM-3 antibodies described herein.

TABLE 8Amino acid sequences of other exemplary anti-TIM-3 antibody moleculesAPE5137 SEQ ID NO: 830 VHEVQLLESGGGLVQPGGSLRLSCAAASGFTFSSYDMSWVRQAPGKGLDWVSTISGGGTYTYYQDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASMD YWGQGTTVTVSSASEQ ID NO: 831 VL DIQMTQSPSSLSASVGDRVTITCRASQSIRRYLNWYHQKPGKAPKLLIYGASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQSHSAPLTFGGGTKVE IKR APE5121SEQ ID NO: 832 VH EVQVLESGGGLVQPGGSLRLYCVASGFTFSGSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKKY YVGPADYWGQGTLVTVSSGSEQ ID NO: 833 VL DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSNNKNYLAWYQHKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYYSSPLTF GGGTKIEVK

Formulations

The anti-TIM-3 antibody molecules described herein can be formulatedinto a formulation (e.g., a dose formulation or dosage form) suitablefor administration (e.g., intravenous administration) to a subject asdescribed herein. The formulation described herein can be a liquidformulation, a lyophilized formulation, or a reconstituted formulation.

In certain embodiments, the formulation is a liquid formulation. In someembodiments, the formulation (e.g., liquid formulation) comprises ananti-TIM-3 antibody molecule (e.g., an anti-TIM-3 antibody moleculedescribed herein) and a buffering agent.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of25 mg/mL to 250 mg/mL, e.g., 50 mg/mL to 200 mg/mL, 60 mg/mL to 180mg/mL, 70 mg/mL to 150 mg/mL, 80 mg/mL to 120 mg/mL, 90 mg/mL to 110mg/mL, 50 mg/mL to 150 mg/mL, 50 mg/mL to 100 mg/mL, 150 mg/mL to 200mg/mL, or 100 mg/mL to 200 mg/mL, e.g., 50 mg/mL, 60 mg/mL, 70 mg/mL, 80mg/mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL,or 150 mg/mL. In certain embodiments, the anti-TIM-3 antibody moleculeis present at a concentration of 80 mg/mL to 120 mg/mL, e.g., 100 mg/mL.

In some embodiments, the formulation (e.g., liquid formulation)comprises a buffering agent comprising histidine (e.g., a histidinebuffer). In certain embodiments, the buffering agent (e.g., histidinebuffer) is present at a concentration of 1 mM to 100 mM, e.g., 2 mM to50 mM, 5 mM to 40 mM, 10 mM to 30 mM, 15 to 25 mM, 5 mM to 40 mM, 5 mMto 30 mM, 5 mM to 20 mM, 5 mM to 10 mM, 40 mM to 50 mM, 30 mM to 50 mM,20 mM to 50 mM, 10 mM to 50 mM, or 5 mM to 50 mM, e.g., 2 mM, 5 mM, 10mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, or 50 mM. In someembodiments, the buffering agent (e.g., histidine buffer) is present ata concentration of 15 mM to 25 mM, e.g., 20 mM. In other embodiments,the buffering agent (e.g., a histidine buffer) or the formulation has apH of 4 to 7, e.g., 5 to 6, e.g., 5, 5.5, or 6. In some embodiments, thebuffering agent (e.g., histidine buffer) or the formulation has a pH of5 to 6, e.g., 5.5. In certain embodiments, the buffering agent comprisesa histidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM)and has a pH of 5 to 6 (e.g., 5.5). In certain embodiments, thebuffering agent comprises histidine and histidine-HCl.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; and a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM), ata pH of 5 to 6 (e.g., 5.5).

In some embodiments, the formulation (e.g., liquid formulation) furthercomprises a carbohydrate. In certain embodiments, the carbohydrate issucrose. In some embodiments, the carbohydrate (e.g., sucrose) ispresent at a concentration of 50 mM to 500 mM, e.g., 100 mM to 400 mM,150 mM to 300 mM, 180 mM to 250 mM, 200 mM to 240 mM, 210 mM to 230 mM,100 mM to 300 mM, 100 mM to 250 mM, 100 mM to 200 mM, 100 mM to 150 mM,300 mM to 400 mM, 200 mM to 400 mM, or 100 mM to 400 mM, e.g., 100 mM,150 mM, 180 mM, 200 mM, 220 mM, 250 mM, 300 mM, 350 mM, or 400 mM. Insome embodiments, the formulation comprises a carbohydrate or sucrosepresent at a concentration of 200 mM to 250 mM, e.g., 220 mM.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM); anda carbohydrate or sucrose present at a concentration of 200 mM to 250mM, e.g., 220 mM, at a pH of 5 to 6 (e.g., 5.5).

In some embodiments, the formulation (e.g., liquid formulation) furthercomprises a surfactant. In certain embodiments, the surfactant ispolysorbate 20. In some embodiments, the surfactant or polysorbate 20)is present at a concentration of 0.005% to 0.1% (w/w), e.g., 0.01% to0.08%, 0.02% to 0.06%, 0.03% to 0.05%, 0.01% to 0.06%, 0.01% to 0.05%,0.01% to 0.03%, 0.06% to 0.08%, 0.04% to 0.08%, or 0.02% to 0.08% (w/w),e.g., 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or0.1% (w/w). In some embodiments, the formulation comprises a surfactantor polysorbate 20 present at a concentration of 0.03% to 0.05%, e.g.,0.04% (w/w).

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM); acarbohydrate or sucrose present at a concentration of 200 mM to 250 mM,e.g., 220 mM; and a surfactant or polysorbate 20 present at aconcentration of 0.03% to 0.05%, e.g., 0.04% (w/w), at a pH of 5 to 6(e.g., 5.5).

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of100 mg/mL; a buffering agent that comprises a histidine buffer (e.g.,histidine/histidine-HCL) at a concentration of 20 mM); a carbohydrate orsucrose present at a concentration of 220 mM; and a surfactant orpolysorbate 20 present at a concentration of 0.04% (w/w), at a pH of 5to 6 (e.g., 5.5).

A formulation described herein can be stored in a container. Thecontainer used for any of the formulations described herein can include,e.g., a vial, and optionally, a stopper, a cap, or both. In certainembodiments, the vial is a glass vial, e.g., a 6R white glass vial. Inother embodiments, the stopper is a rubber stopper, e.g., a grey rubberstopper. In other embodiments, the cap is a flip-off cap, e.g., analuminum flip-off cap. In some embodiments, the container comprises a 6Rwhite glass vial, a grey rubber stopper, and an aluminum flip-off cap.In some embodiments, the container (e.g., vial) is for a single-usecontainer. In certain embodiments, 25 mg/mL to 250 mg/mL, e.g., 50 mg/mLto 200 mg/mL, 60 mg/mL to 180 mg/mL, 70 mg/mL to 150 mg/mL, 80 mg/mL to120 mg/mL, 90 mg/mL to 110 mg/mL, 50 mg/mL to 150 mg/mL, 50 mg/mL to 100mg/mL, 150 mg/mL to 200 mg/mL, or 100 mg/mL to 200 mg/mL, e.g., 50mg/mL, 60 mg/mL, 70 mg/mL, 80 mg/mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120mg/mL, 130 mg/mL, 140 mg/mL, or 150 mg/mL, of the anti-TIM-3 antibodymolecule, is present in the container (e.g., vial).

In another aspect, the disclosure features therapeutic kits that includethe anti-TIM-3 antibody molecules, compositions, or formulationsdescribed herein, and instructions for use, e.g., in accordance withdosage regimens described herein.

Hypomethylating Agents

In certain embodiments, the combination described herein includes ahypomethylating agent. Hypomethylating agents are also known as HMAs ordemethylating agents, which inhibits DNA methylation. In certainembodiments, the hypomethylating agent blocks the activity of DNAmethyltransferase. In certain embodiments, the hypomethylating agentcomprises azacitidine, decitabine, CC-486 (Bristol Meyers Squibb), orASTX727 (Astex).

In some embodiments, the combination described herein to treat MDS(e.g., an intermediate MDS, a high risk MDS, or a very high risk MDS) ora CMML (e.g., a CMML-1 or a CMML-2), comprises a TIM-3 inhibitordescribed herein, e.g., MBG453) administered intravenously at a dose of600 mg to 1000 mg (e.g., 800 mg), e.g., over 30 minutes, e.g., on day 8of each 28 day cycle; and a hypomethylating agent described herein(e.g., azacitidine) administered intravenously or subcutaneously at adose of 50 mg/m² to 100 mg/m² (e.g., 75 mg/m²), e.g., on sevenconsecutive days, e.g., days 1, 2, 3, 4, 5, 6, and 7, of a 28 day cycle.In other embodiments described herein to treat MDS (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS) or a CMML(e.g., a CMML-1 or a CMML-2), comprises a TIM-3 inhibitor describedherein, e.g., MBG453) administered intravenously at a dose of 600 mg to1000 mg (e.g., 800 mg), e.g., over 30 minutes on day 8 of each 28 daycycle; and a hypomethylating agent described herein (e.g., azacitidine)administered intravenously or subcutaneously at a dose of 50 mg/m² to100 mg/m² (e.g., 75 mg/m²), e.g., on days 1, 2, 3, 4, and 5, and days 8and 9 of a 28 day cycle. In some embodiments, the TIM-3 inhibitor (e.g.,MBG453), and the hypomethylating agent are administered on the same day.In some embodiments, the TIM-3 inhibitor (e.g., MBG453) is administeredafter administration of the hypomethylating agent (e.g., azacitidine)has completed. In some embodiments, the TIM-3 inhibitor is administeredabout 30 minutes to about four hours (e.g., about one hour afteradministration of the hypomethylating agent (e.g., azacitidine) hascompleted.

Exemplary Hypomethylating Agents

In some embodiments, the hypomethylating agent comprises azacitidine.Azacitidine is also known as 5-AC, 5-azacytidine, azacytidine,ladakamycin, 5-AZC, AZA-CR, U-18496,4-amino-1-beta-D-ribofuranosyl-1,3,5-triazin-2(1H)-one,4-amino-1-[(2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-1,3,5-triazin-2-one,or VIDAZA®. Azacitidine has the following structural formula:

or a pharmaceutically acceptable salt thereof.

Azacitidine is a pyrimidine nucleoside analogue of cytidine withantineoplastic activity. Azacitidine is incorporated into DNA, where itreversibly inhibits DNA methyltransferase, thereby blocking DNAmethylation. Hypomethylation of DNA by azacitidine can activate tumorsuppressor genes silenced by hypermethylation, resulting in an antitumoreffect. Azacitidine can also be incorporated into RNA, therebydisrupting normal RNA function and impairing tRNA cytosinemethyltransferase activity.

In some embodiments, azacitidine is administered at a dose of about 25mg/m² to about 150 mg/m², e.g., about 50 mg/m² to about 100 mg/m², about70 mg/m² to about 80 mg/m², about 50 mg/m² to about 75 mg/m², about 75mg/m² to about 125 mg/m², about 50 mg/m², about 75 mg/m², about 100mg/m², about 125 mg/m², or about 150 mg/m². In some embodiments,azacitidine is administered once a day. In some embodiments, azacitidineis administered intravenously. In other embodiments, azacitidine isadministered subcutaneously. In some embodiments, azacitidine isadministered at a dose of about 50 mg/m² to about 100 mg/m² (e.g., about75 mg/m²), e.g., for about 5-7 consecutive days, e.g., in a 28-daycycle. For example, azacitidine can be administered at a dose of about75 mg/m² for seven consecutive days on days 1-7 of a 28-day cycle. Asanother example, azacitidine can be administered at a dose of about 75mg/m² for five consecutive days on days 1-5 of a 28-day cycle, followedby a two-day break, then two consecutive days on days 8-9.

Other Exemplary Hypomethylating Agents

In some embodiments, the hypomethylating agent comprises decitabine,CC-486, or ASTX727.

Decitabine is also known as 5-aza-dCyd, deoxyazacytidine, dezocitidine,SAZA, DAC, 2′-deoxy-5-azacytidine,4-amino-1-(2-deoxy-beta-D-erythro-pentofuranosyl)-1,3,5-triazin-2(1H)-one,5-aza-2′-deoxycytidine, 5-aza-2-deoxycytidine, 5-azadeoxycytidine, orDACOGEN®. Decitabine has the following structural formula:

or a pharmaceutically acceptable salt thereof.

Decitabine is a cytidine antimetabolite analogue with potentialantineoplastic activity. Decitabine incorporates into DNA and inhibitsDNA methyltransferase, resulting in hypomethylation of DNA andintra-S-phase arrest of DNA replication.

In some embodiments, decitabine is administered at a dose of about 5mg/m² to about 50 mg/m², e.g., about 10 mg/m² to about 40 mg/m², about20 mg/m² to about 30 mg/m², about 5 mg/m² to about 40 mg/m², about 5mg/m² to about 30 mg/m², about 5 mg/m² to about 20 mg/m², about 5 mg/m²to about 10 mg/m², about 10 mg/m² to about 50 mg/m², about 20 mg/m² toabout 50 mg/m², about 30 mg/m² to about 50 mg/m², about 40 mg/m² toabout 50 mg/m², about 10 mg/m² to about 20 mg/m², about 15 mg/m² toabout 25 mg/m², about 5 mg/m², about 10 mg/m², about 15 mg/m², about 20mg/m², about 25 mg/m², about 30 mg/m², about 35 mg/m², about 40 mg/m²,about 45 mg/m², or about 50 mg/m². In some embodiments, decitabine isadministered intravenously. In certain embodiments, decitabine isadministered according a three-day regimen, e.g., administered at a doseof about 10 mg/m² to about 20 mg/m² (e.g., 15 mg/m²) by continuousintravenous infusion over about 3 hours repeated every 8 hours for 3days (repeat cycles every 6 weeks, e.g., for a minimum of 4 cycles). Inother embodiments, decitabine is administered according to a five-dayregimen, e.g., administered at a dose of about 10 mg/m² to about 20mg/m² (e.g., 15 mg/m²) by continuous intravenous infusion over about 1hour daily for 5 days (repeat cycles every 4 weeks, e.g., for a minimumof 4 cycles).

In some embodiments, the hypomethylating agent comprises an oralazacitidine (e.g., CC-486). In some embodiments, the hypomethylatingagent comprises CC-486. CC-486 is an orally bioavailable formulation ofazacitidine, a pyrimidine nucleoside analogue of cytidine, withantineoplastic activity. Upon oral administration, azacitidine is takenup by cells and metabolized to 5-azadeoxycitidine triphosphate. Theincorporation of 5-azadeoxycitidine triphosphate into DNA reversiblyinhibits DNA methyltransferase, and blocks DNA methylation.Hypomethylation of DNA by azacitidine can re-activate tumor suppressorgenes previously silenced by hypermethylation, resulting in an antitumoreffect. The incorporation of 5-azacitidine triphosphate into RNA candisrupt normal RNA function and impairs tRNA(cytosine-5)-methyltransferase activity, resulting in an inhibition ofRNA and protein synthesis. CC-486 is described, e.g., in Laille et al. JClin Pharmacol. 2014; 54(6):630-639; Mesia et al. European Journal ofCancer 2019 123:138-154. Oral formulations of cytidine analogs are alsodescribed, e.g., in PCT Publication No. WO 2009/139888 and U.S. Pat. No.8,846,628. In some embodiments, CC-486 is administered orally. In someembodiments, CC-486 is administered on once daily. In some embodiments,CC-486 is administered at a dose of about 200 mg to about 500 mg (e.g.,300 mg). In some embodiments, CC-486 is administered on 5-15 consecutivedays (e.g., days 1-14) of, e.g., a 21 day or 28 day cycle. In someembodiments, CC-486 is administered once a day.

In some embodiments, the hypomethylating agent comprises a CDA inhibitor(e.g., cedazuridine/decitabine combination agent (e.g., ASTX727)). Insome embodiments, the hypomethylating agent comprises ASTX727. ASTX727is an orally available combination agent comprising the cytidinedeaminase (CDA) inhibitor cedazuridine (also known as E7727) and thecytidine antimetabolite decitabine, with antineoplastic activity. Uponoral administration of ASTX727, the CDA inhibitor E7727 binds to andinhibits CDA, an enzyme primarily found in the gastrointestinal (GI)tract and liver that catalyzes the deamination of cytidine and cytidineanalogs. This can prevent the breakdown of decitabine, increasing itsbioavailability and efficacy while decreasing GI toxicity due to theadministration of lower doses of decitabine. Decitabine exerts itsantineoplastic activity through the incorporation of its triphosphateform into DNA, which inhibits DNA methyltransferase and results inhypomethylation of DNA. This can interfere with DNA replication anddecreases tumor cell growth. ASTX727 is disclosed in e.g.,Montalaban-Bravo et al. Current Opinions in Hematology 201825(2):146-153. In some embodiments, ASTX727 comprises cedazuridine,e.g., about 50-150 mg (e.g., about 100 mg), and decitabine, e.g., about300-400 mg (e.g., 345 mg). In some embodiments, ASTX727 is administeredorally. In some embodiments, ASTX727 is administered on 5-15 consecutivedays (e.g., days 1-5) of, e.g., a 28 day cycle. In some embodiments,ASTX727 is administered once a day.

Cytarabine

In some embodiments, the combination described herein includescytarabine. Cytarabine is also known as cytosine arabinoside or4-amino-1-[(2R,3S,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one.Cytarabine has the following structural formula:

or a pharmaceutically acceptable salt thereof.

Cytarabine is a cytidine antimetabolite analogue with a modified sugarmoiety (arabinose in place of ribose). Cytarabine is converted to atriphosphate form which competes with cytidine for incorporation intoDNA. Due to the arabinose sugar, the rotation of the DNA molecule issterically hindered and DNA replication ceases. Cytarabine alsointerferes with DNA polymerase.

In some embodiments, cytarabine is administered at about 5 mg/m² toabout 75 mg/m², e.g., 30 mg/m². In some embodiments, cytarabine isadministered about 100 mg/m² to about 400 mg/m², e.g., 100 mg/m². Insome embodiments, cytarabine is administered by intravenous infusion orinjection, subcutaneously, or intrathecally. In some embodiments,cytarabine is administered at a dose of 100 mg/m²/day by continuous IVinfusion or 100 mg/m² intravenously every 12 hours. In some embodiments,cytarabine is administered for 7 days (e.g. on days 1 to 7). In someembodiments, cytarabine is administered intrathecally at a dose rangingfrom 5 to 75 mg/m² of body surface area. In some embodiments, cytarabineis intrathecally administered from once every 4 days to once a day for 4days. In some embodiments, cytarabine is administered at a dose of 30mg/m² every 4 days.

Further Combinations

The combinations described herein can further comprises one or moreother therapeutic agents, procedures or modalities.

In one embodiment, the methods described herein include administering tothe subject a combination comprising a TIM-3 inhibitor described hereinand a hypomethylating agent described herein, in combination with atherapeutic agent, procedure, or modality, in an amount effective totreat or prevent a disorder described herein. In certain embodiments,the combination is administered or used in accordance with a dosageregimen described herein. In other embodiments, the combination isadministered or used as a composition or formulation described herein.

The TIM-3 inhibitor, hypomethylating agent, and the therapeutic agent,procedure, or modality can be administered or used simultaneously orsequentially in any order. Any combination and sequence of the TIM-3inhibitor, hypomethylating agent, and the therapeutic agent, procedure,or modality (e.g., as described herein) can be used. The TIM-3inhibitor, hypomethylating agent, and/or the therapeutic agent,procedure or modality can be administered or used during periods ofactive disorder, or during a period of remission or less active disease.The TIM-3 inhibitor, or hypomethylating agent can be administeredbefore, concurrently with, or after the treatment with the therapeuticagent, procedure or modality.

In certain embodiments, the combination described herein can beadministered with one or more of other antibody molecules, chemotherapy,other anti-cancer therapy (e.g., targeted anti-cancer therapies, genetherapy, viral therapy, RNA therapy bone marrow transplantation,nanotherapy, or oncolytic drugs), cytotoxic agents, immune-basedtherapies (e.g., cytokines or cell-based immune therapies), surgicalprocedures (e.g., lumpectomy or mastectomy) or radiation procedures, ora combination of any of the foregoing. The additional therapy may be inthe form of adjuvant or neoadjuvant therapy. In some embodiments, theadditional therapy is an enzymatic inhibitor (e.g., a small moleculeenzymatic inhibitor) or a metastatic inhibitor. Exemplary cytotoxicagents that can be administered in combination with includeantimicrotubule agents, topoisomerase inhibitors, antimetabolites,mitotic inhibitors, alkylating agents, anthracyclines, vinca alkaloids,intercalating agents, agents capable of interfering with a signaltransduction pathway, agents that promote apoptosis, proteasomeinhibitors, and radiation (e.g., local or whole-body irradiation (e.g.,gamma irradiation). In other embodiments, the additional therapy issurgery or radiation, or a combination thereof. In other embodiments,the additional therapy is a therapy targeting one or more ofPI3K/AKT/mTOR pathway, an HSP90 inhibitor, or a tubulin inhibitor.

Alternatively, or in combination with the aforesaid combinations, thecombination described herein can be administered or used with, one ormore of an inhibitor of CD47, CD70, NEDD8, CDK9, MDM2, FLT3, or KITand/or an activator of p53. In some embodiments, the TIM-3 inhibitor isadministered with an inhibitor of CD47, CD70, NEDD8, CDK9, MDM2, FLT3,or KIT and/or an activator of p53. In some embodiments, the TIM-3inhibitor is administered with a hypomethylating agent, e.g., ahypomethylating agent described herein, further in combination with aninhibitor of CD47, CD70, NEDD8, CDK9, MDM2, FLT3, or KIT and/or anactivator of p53.

In some embodiments, the TIM-3 inhibitor is administered with ahypomethylating agent, e.g., a hypomethylating agent described herein,further in combination with an inhibitor of CD47, CD70, NEDD8, CDK9,MDM2, FLT3, or KIT and/or an activator of p53 to treat MDS (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS).

In some embodiments, the TIM-3 inhibitor is administered with ahypomethylating agent, e.g., a hypomethylating agent described herein,further in combination with an inhibitor of CD47, CD70, NEDD8, CDK9,MDM2, FLT3, or KIT and/or an activator of p53 to treat a CMML (e.g., aCMML-1 or a CMML-2).

Alternatively, or in combination with the aforesaid combinations, thecombination described herein can be administered or used with, one ormore of: an immunomodulator (e.g., an activator of a costimulatorymolecule or an inhibitor of an inhibitory molecule, e.g., an immunecheckpoint molecule); a vaccine, e.g., a therapeutic cancer vaccine; orother forms of cellular immunotherapy.

In certain embodiments, the combination described herein is administeredor used in with a modulator of a costimulatory molecule or an inhibitorymolecule, e.g., a co-inhibitory ligand or receptor.

In one embodiment, the compounds and combinations described herein areadministered or used with a modulator, e.g., agonist, of a costimulatorymolecule. In one embodiment, the agonist of the costimulatory moleculeis chosen from an agonist (e.g., an agonistic antibody orantigen-binding fragment thereof, or a soluble fusion) of OX40, CD2,CD27, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), 4-1BB (CD137),GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160,B7-H3 or CD83 ligand.

In another embodiment, the compounds and/or combinations describedherein are administered or used in combination with a GITR agonist,e.g., an anti-GITR antibody molecule.

In one embodiment, the compounds and/or combinations described hereinare administered or used in combination with an inhibitor of aninhibitory (or immune checkpoint) molecule chosen from PD-L1, PD-L2,CTLA-4, TIM-3, LAG-3, CEACAM (e.g., CEACAM-1, CEACAM-3, and/orCEACAM-5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 and/or TGF beta. In oneembodiment, the inhibitor is a soluble ligand (e.g., a CTLA-4-Ig), or anantibody or antibody fragment that binds to PD-1, LAG-3, PD-L1, PD-L2,or CTLA-4.

In another embodiment, the compounds and/or combinations describedherein are administered or used in combination with a PD-1 inhibitor,e.g., an anti-PD-1 antibody molecule. In another embodiment, theanti-TIM-3 antibody molecule described herein is administered or used incombination with a LAG-3 inhibitor, e.g., an anti-LAG-3 antibodymolecule. In another embodiment, the anti-TIM-3 antibody moleculedescribed herein is administered or used in combination with a PD-L1inhibitor, e.g., an anti-PD-L1 antibody molecule.

In another embodiment, the compounds and/or combinations describedherein are administered or used in combination with a PD-1 inhibitor(e.g., an anti-PD-1 antibody molecule) and a LAG-3 inhibitor (e.g., ananti-LAG-3 antibody molecule). In another embodiment, the anti-TIM-3antibody molecule described herein is administered or used incombination with a PD-1 inhibitor (e.g., an anti-PD-1 antibody molecule)and a PD-L1 inhibitor (e.g., an anti-PD-L1 antibody molecule). Inanother embodiment, the anti-TIM-3 antibody molecule described herein isadministered or used in combination with a LAG-3 inhibitor (e.g., ananti-LAG-3 antibody molecule) and a PD-L1 inhibitor (e.g., an anti-PD-L1antibody molecule).

In another embodiment, the compounds and/or combinations describedherein are administered or used in combination with a CEACAM inhibitor(e.g., CEACAM-1, CEACAM-3, and/or CEACAM-5 inhibitor), e.g., ananti-CEACAM antibody molecule. In another embodiment, the anti-TIM-3antibody molecule is administered or used in combination with a CEACAM-1inhibitor, e.g., an anti-CEACAM-1 antibody molecule. In anotherembodiment, the anti-TIM-3 antibody molecule is administered or used incombination with a CEACAM-3 inhibitor, e.g., an anti-CEACAM-3 antibodymolecule. In another embodiment, the anti-PD-1 antibody molecule isadministered or used in combination with a CEACAM-5 inhibitor, e.g., ananti-CEACAM-5 antibody molecule.

The combination of antibody molecules disclosed herein can beadministered separately, e.g., as separate antibody molecules, orlinked, e.g., as a bispecific or trispecific antibody molecule. In oneembodiment, a bispecific antibody that includes an anti-TIM-3 antibodymolecule and an anti-PD-1, anti-CEACAM (e.g., anti-CEACAM-1, CEACAM-3,and/or anti-CEACAM-5), anti-PD-L1, or anti-LAG-3 antibody molecule, isadministered. In certain embodiments, the combination of antibodiesdisclosed herein is used to treat a cancer, e.g., a cancer as describedherein (e.g., a solid tumor or a hematologic malignancy).

CD47 Inhibitor

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a CD47 inhibitor. In someembodiments, the CD47 inhibitor is magrolimab. In some embodiments,these combinations are used to treat the cancer indications disclosedherein, including the hematologic indications disclosed herein,including an MDS (e.g., an intermediate MDS, a high risk MDS, or a veryhigh risk MDS). In some embodiments, these combinations are used totreat the cancer indications disclosed herein, including the hematologicindications disclosed herein, including a CMML (e.g., a CMML-1 or aCMML-2).

Exemplary CD47 Inhibitor

In some embodiments, the CD47 inhibitor is an anti-CD47 antibodymolecule. In some embodiments, the anti-CD47 antibody comprisesmagrolimab. Magrolimab is also known as ONO-7913, 5F9, or Hu5F9-G4.Magrolimab selectively binds to CD47 expressed on tumor cells and blocksthe interaction of CD47 with its ligand signal regulatory protein alpha(SIRPa), a protein expressed on phagocytic cells. This typicallyprevents CD47/SIRPa-mediated signaling, allows the activation ofmacrophages, through the induction of pro-phagocytic signaling mediatedby calreticulin, which is specifically expressed on the surface of tumorcells, and results in specific tumor cell phagocytosis. In addition,blocking CD47 signaling generally activates an anti-tumor T-lymphocyteimmune response and T-mediated cell killing Magrolimab is disclosed,e.g., in Sallaman et al. Blood 2019 134(Supplement_1):569.

In some embodiments, magrolimab is administered intravenously. In someembodiments, magrolimab is administered on days 1, 4, 8, 11, 15, and 22of cycle 1 (e.g., a 28 day cycle), days 1, 8, 15, and 22 of cycle 2(e.g., a 28 day cycle), and days 1 and 15 of cycle 3 (e.g., a 28 daycycle) and subsequent cycles. In some embodiments, magrolimab isadministered at least twice weekly, each week of, e.g., a 28 day cycle.In some embodiments, magrolimab is administered in a dose-escalationregimen. In some embodiments, magrolimab is administered at 1-30 mg/kg,e.g., 1-30 mg/kg per week.

Other CD47 Inhibitors

In some embodiments, the CD47 inhibitor is an inhibitor chosen fromB6H12.2, CC-90002, C47B157, C47B161, C47B222, SRF231, ALX148, W6/32,4N1K, 4N1, TTI-621, TTI-622, PKHB1, SEN177, MiR-708, and MiR-155. Insome embodiments, the CD47 inhibitor is a bispecific antibody.

In some embodiments, the CD47 inhibitor is B6H12.2. B6H12.2 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. B6H12.2 is ahumanized anti-CD74-IgG4 antibody that binds to CD47 expressed on tumorcells and blocks the interaction of CD47 with its ligand signalregulatory protein alpha (SIRPa).

In some embodiments, the CD47 inhibitor is CC-90002. CC-90002 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. CC-90002 is amonoclonal antibody targeting the human cell surface antigen CD47, withpotential phagocytosis-inducing and antineoplastic activities. Uponadministration, anti-CD47 monoclonal antibody CC-90002 selectively bindsto CD47 expressed on tumor cells and blocks the interaction of CD47 withsignal regulatory protein alpha (SIRPa), a protein expressed onphagocytic cells. This prevents CD47/SIRPa-mediated signaling andabrogates the CD47/SIRPa-mediated inhibition of phagocytosis. Thisinduces pro-phagocytic signaling mediated by the binding of calreticulin(CRT), which is specifically expressed on the surface of tumor cells, tolow-density lipoprotein (LDL) receptor-related protein (LRP), expressedon macrophages. This results in macrophage activation and the specificphagocytosis of tumor cells. In addition, blocking CD47 signalingactivates both an anti-tumor T-lymphocyte immune response and Tcell-mediated killing of CD47-expressing tumor cells. In someembodiments, CC-90002 is administered intravenously. In someembodiments, CC-90002 is administered intravenously on a 28-day cycle.

In some embodiments, the CD47 inhibitor is C47B157, C47B161, or C47B222.C47B157, C47B161, and C47B222 are disclosed, e.g., in Eladl et al.Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. C47B157, C47B161, andC47B222 are humanized anti-CD74-IgG1 antibodies that bind to CD47expressed on tumor cells and blocks the interaction of CD47 with itsligand signal regulatory protein alpha (SIRPa).

In some embodiments, the CD47 inhibitor is SRF231. SRF231 is disclosed,e.g., in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. SRF231 is a human monoclonalantibody targeting the human cell surface antigen CD47, with potentialphagocytosis-inducing and antineoplastic activities. Uponadministration, anti-CD47 monoclonal antibody SRF231 selectively bindsto CD47 on tumor cells and blocks the interaction of CD47 with signalregulatory protein alpha (SIRPalpha), an inhibitory protein expressed onmacrophages. This prevents CD47/SIRPalpha-mediated signaling andabrogates the CD47/SIRPa-mediated inhibition of phagocytosis. Thisinduces pro-phagocytic signaling mediated by the binding of calreticulin(CRT), which is specifically expressed on the surface of tumor cells, tolow-density lipoprotein (LDL) receptor-related protein (LRP), expressedon macrophages. This results in macrophage activation and the specificphagocytosis of tumor cells. In addition, blocking CD47 signalingactivates both an anti-tumor T-lymphocyte immune response andT-cell-mediated killing of CD47-expressing tumor cells.

In some embodiments, the CD47 inhibitor is ALX148. ALX148 is disclosed,e.g., in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. ALX148 is a CD47 antagonist.It is a variant of signal regulatory protein alpha (SIRPa) thatantagonizes the human cell surface antigen CD47, with potentialphagocytosis-inducing, immunostimulating and antineoplastic activities.Upon administration, ALX148 binds to CD47 expressed on tumor cells andprevents the interaction of CD47 with its ligand SIRPa, a proteinexpressed on phagocytic cells. This prevents CD47/SIRPa-mediatedsignaling and abrogates the CD47/SIRPa-mediated inhibition ofphagocytosis. This induces pro-phagocytic signaling mediated by thebinding of the pro-phagocytic signaling protein calreticulin (CRT),which is specifically expressed on the surface of tumor cells, tolow-density lipoprotein (LDL) receptor-related protein (LRP), expressedon macrophages. This results in macrophage activation and the specificphagocytosis of tumor cells. In addition, blocking CD47 signalingactivates both an anti-tumor cytotoxic T-lymphocyte (CTL) immuneresponse and T-cell-mediated killing of CD47-expressing tumor cells. Insome embodiments, ALX148 is administered intravenously. In someembodiments, ALX148 is administered at least once a week. In someembodiments, ALX148 is administered at least twice a week.

In some embodiments, the CD47 inhibitor is W6/32. W6/32 is disclosed,e.g., in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. W6/32 is an anti-CD47antibody that targets CD47-MHC-1.

In some embodiments, the CD47 inhibitor is 4N1K or 4N1. 4N1K and 4N1 aredisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. 4N1K and 4N1 areCD47-SIRPa Peptide agonists.

In some embodiments, the CD47 inhibitor is TTI-621. TTI-621 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. TTI-621 is also knownas SIRPa-IgG1 Fc. TTI-621 is a soluble recombinant antibody-like fusionprotein composed of the N-terminal CD47 binding domain of humansignal-regulatory protein alpha (SIRPa) linked to the Fc domain of humanimmunoglobulin G1 (IgG1), with potential immune checkpoint inhibitoryand antineoplastic activities. Upon administration, the SIRPa-Fc fusionprotein TTI-621 selectively targets and binds to CD47 expressed on tumorcells and blocks the interaction of CD47 with endogenous SIRPa, a cellsurface protein expressed on macrophages. This preventsCD47/SIRPa-mediated signaling and abrogates the CD47/SIRPa-mediatedinhibition of macrophage activation and phagocytosis of cancer cells.This induces pro-phagocytic signaling mediated by the binding ofcalreticulin (CRT), which is specifically expressed on the surface oftumor cells, to low-density lipoprotein (LDL) receptor-related protein-1(LRP-1), expressed on macrophages, and results in macrophage activationand the specific phagocytosis of tumor cells. In some embodiments,TTI-621 is administered intratumorally.

In some embodiments, the CD47 inhibitor is TTI-622. TTI-622 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. TTI-622 is also knownas SIRPa-IgG1 Fc. TTI-622 is a soluble recombinant antibody-like fusionprotein composed of the N-terminal CD47 binding domain of humansignal-regulatory protein alpha (SIRPa; CD172a) linked to an Fc domainderived from human immunoglobulin G subtype 4 (IgG4), with potentialimmune checkpoint inhibitory, phagocytosis-inducing and antineoplasticactivities. Upon administration, the SIRPa-IgG4-Fc fusion proteinTTI-622 selectively targets and binds to CD47 expressed on tumor cellsand blocks the interaction of CD47 with endogenous SIRPa, a cell surfaceprotein expressed on macrophages. This prevents CD47/SIRPa-mediatedsignaling and abrogates the CD47/SIRPa-mediated inhibition of macrophageactivation. This induces pro-phagocytic signaling resulting from thebinding of calreticulin (CRT), which is specifically expressed on thesurface of tumor cells, to low-density lipoprotein (LDL)receptor-related protein-1 (LRP-1) expressed on macrophages, and resultsin macrophage activation and the specific phagocytosis of tumor cells.

In some embodiments, the CD47 inhibitor is PKHB1. PKHB1 is disclosed,e.g., in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. PKHB1 is a CD47 peptideagonist that binds CD47 and blocks the interaction with SIRPα.

In some embodiments, the CD47 inhibitor is SEN177. SEN177 is disclosed,e.g., in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1. SEN177 is an antibody thattargets QPCTL in CD47.

In some embodiments, the CD47 inhibitor is MiR-708. MiR-708 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. MiR-708 is a miRNAthat targets CD47 and blocks the interaction with SIRPα.

In some embodiments, the CD47 inhibitor is MiR-155. MiR-155 isdisclosed, e.g., in Eladl et al. Journal of Hematology & Oncology 202013(96) https://doi.org/10.1186/s13045-020-00930-1. MiR-155 is a miRNAthat targets CD47 and blocks the interaction with SIRPα.

In some embodiments, the CD47 inhibitor is an anti-CD74, anti-PD-L1bispecific antibody or an anti-CD47, anti-CD20 bispecific antibody, asdisclosed in Eladl et al. Journal of Hematology & Oncology 2020 13(96)https://doi.org/10.1186/s13045-020-00930-1.

In some embodiments, the CD74 inhibitor is LicMAB as disclosed in, e.g.,Ponce et al. Oncotarget 2017 8(7):11284-11301.

CD70 Inhibitor

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a CD70 inhibitor. In someembodiments, the CD70 inhibitor is cusatuzumab. In some embodiments,these combinations are used to treat the cancer indications disclosedherein, including the hematologic indications disclosed herein,including an MDS (e.g., an intermediate MDS, a high risk MDS, or a veryhigh risk MDS). In some embodiments, these combinations are used totreat the cancer indications disclosed herein, including the hematologicindications disclosed herein, including a CMML (e.g., a CMML-1 or aCMML-2).

Exemplary CD70 Inhibitor

In some embodiments, the CD70 inhibitor is an anti-CD70 antibodymolecule. In some embodiments, the anti-CD70 antibody comprisescusatuzumab. Cusatuzumab is also known as ARGX-110 or JNJ-74494550.Cusatuzumab selectively binds to, and neutralizes the activity of CD70,which may also induce an antibody-dependent cellular cytotoxicity (ADCC)response against CD70-expressing tumor cells. Cusatuzumab is disclosed,e.g., in Riether et al. Nature Medicine 2020 26:1459-1467.

In some embodiments, cusatuzumab is administered intravenously. In someembodiments, cusatuzumab is administered subcutaneously. In someembodiments, cusatuzumab is administered at 1-20 mg/kg, e.g., 1 mg/kg, 3mg/kg, 10 mg/kg, or 20 mg/kg. In some embodiments, cusatuzumab isadministered once every two weeks. In some embodiments, cusatuzumab isadministered at 10 mg/kg once every two weeks. In some embodiments,cusatuzumab is administered at 20 mg/kg once every two weeks. In someembodiments, cusatuzumab is administered on day 3 and day 17 of, e.g., a28 day cycle.

p53 Activator

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a p53 activator. In someembodiments, the p53 activator is APR-246. In some embodiments, thesecombinations are used to treat the cancer indications disclosed herein,including the hematologic indications disclosed herein, including an MDS(e.g., an intermediate MDS, a high risk MDS, or a very high risk MDS).In some embodiments, these combinations are used to treat the cancerindications disclosed herein, including the hematologic indicationsdisclosed herein, including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary p53 Activator

In some embodiments, the p53 activator is APR-246. APR-246 is amethylated derivative and structural analog of PRIMA-1 (p53re-activation and induction of massive apoptosis). APR-246 is also knownas Eprenetapopt, PRIMA-1MET. APR-246 covalently modifies the core domainof mutated forms of cellular tumor p53 through the alkylation of thiolgroups. These modifications restore both the wild-type conformation andfunction to mutant p53, which reconstitutes endogenous p53 activity,leading to cell cycle arrest and apoptosis in tumor cells. APR-246 isdisclosed, e.g., in Zhang et al. Cell Death and Disease 2018 9(439).

In some embodiments, APR-246 is administered on days 1˜4 of, e.g., a28-day cycle, e.g., for 12 cycles. In some embodiments, APR-246 isadministered at 4-5 g, e.g., 4.5 g, each day.

NEDD8 Inhibitor

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a NEDD8 inhibitor. In someembodiments, the NEDD8 inhibitor is an inhibitor of NEDD8 activatingenzyme (NAE). In some embodiments, the NEDD8 inhibitor is pevonedistat.In some embodiments, these combinations are used to treat the cancerindications disclosed herein, including the hematologic indicationsdisclosed herein, including an MDS (e.g., an intermediate MDS, a highrisk MDS, or a very high risk MDS). In some embodiments, thesecombinations are used to treat the cancer indications disclosed herein,including the hematologic indications disclosed herein, including a CMML(e.g., a CMML-1 or a CMML-2).

Exemplary NEDD8 Inhibitor

In some embodiments, the NEDD8 inhibitor is a small molecule inhibitor.In some embodiments, the NEDD8 inhibitor is pevonedistat. Pevonedistatis also known as TAK-924, NAE inhibitor MLN4924, Nedd8-activating enzymeinhibitor MLN4924, MLN4924, or((1S,2S,4R)-4-(4-((1S)-2,3-Dihydro-1H-inden-1-ylamino)-7H-pyrrolo(2,3-d)pyrimidin-7-yl)hydroxycyclopentyl)methyl sulphamate. Pevonedistat binds to and inhibitsNAE, which may result in the inhibition of tumor cell proliferation andsurvival. NAE activates Nedd8 (Neural precursor cell expressed,developmentally down-regulated 8), a ubiquitin-like (UBL) protein thatmodifies cellular targets in a pathway that is parallel to but distinctfrom the ubiquitin-proteasome pathway (UPP). Pevonedistat is disclosed,e.g., in Swords et al. Blood (2018) 131(13)1415-1424.

In some embodiments, pevonedistat is administered intravenously. In someembodiments, pevonedistat is administered at 10-50 mg/m², e.g., 10mg/m², 20 mg/m², 25 mg/m², 30 mg/m², or 50 mg/m². In some embodiments,pevonedistat is administered on days 1, 3, and 5 of, e.g., a 28-daycycle, for, e.g., up to 16 cycles. In some embodiments, pevonedistat isadministered using fixed dosing. In some embodiments, pevonedistat isadministered in a ramp-up dosing schedule. In some embodiments,pevonedistat is administered at 25 mg/m² on day 1 and 50 mg/m² on day 8of, e.g., each 28 day cycle.

CDK9 Inhibitors

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a cyclin dependent kinaseinhibitor. In some embodiments, the combination described herein isfurther administered in combination with a CDK9 inhibitor. In someembodiments, the CDK9 inhibitor is chosen from alvocidib or alvocidibprodrug TP-1287. In some embodiments, these combinations are used totreat the cancer indications disclosed herein, including the hematologicindications disclosed herein, including an MDS (e.g., an intermediateMDS, a high risk MDS, or a very high risk MDS). In some embodiments,these combinations are used to treat the cancer indications disclosedherein, including the hematologic indications disclosed herein,including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary CDK9 Inhibitor

In some embodiments, the CDK9 inhibitor is Alvocidib. Alvocidib is alsoknown as flavopiridol, FLAVO, HMR 1275, L-868275, or(−)-2-(2-chlorophenyl)-5,7-dihydroxy-8-[(3R,4S)-3-hydroxy-1-methyl-4-piperidinyl]-4H-1-benzopyran-4-onehydrochloride. Alvocidib is a synthetic N-methylpiperidinyl chlorophenylflavone compound. As an inhibitor of cyclin-dependent kinase, alvocidibinduces cell cycle arrest by preventing phosphorylation ofcyclin-dependent kinases (CDKs) and by down-regulating cyclin D1 and D3expression, resulting in G1 cell cycle arrest and apoptosis. This agentis also a competitive inhibitor of adenosine triphosphate activity.Alvocidib is disclosed, e.g., in Gupta et al. Cancer Sensitizing Agentsfor Chemotherapy 2019: pp. 125-149.

In some embodiments, alvocidib is administered intravenously. In someembodiments, alvocidib is administered on days 1, 2, and/or 3 of, e.g.,a 28 day cycle. In some embodiments, alvocidib is administered usingfixed dosing. In some embodiments, alvocidib is administered in aramp-up dosing schedule. In some embodiments, alvocidib is administeredfor 4-weeks, followed by a 2 week rest period, for, e.g., up to amaximum of 6 cycles (e.g., a 28 day cycle). In some embodiments,alvocidib is administered at 30-50 mg/m², e.g., 30 mg/m² or 50 mg/m². Insome embodiments, alvocidib is administered at 30 mg/m² as a 30-minuteintravenous (IV) infusion followed by 30 mg/m² as a 4-hour continuousinfusion. In some embodiments, alvocidib is administered at 30 mg/m2over 30 minutes followed by 50 mg/m2 over 4 hours. In some embodiments,alvocidib is administered at a first dose of 30 mg/m² as a 30-minuteintravenous (IV) infusion followed by 30 mg/m² as a 4-hour continuousinfusion, and one or more subsequent doses of 30 mg/m2 over 30 minutesfollowed by 50 mg/m2 over 4 hours.

Other CDK9 Inhibitor

In some embodiments, the CDK9 inhibitor is TP-1287. TP-1287 is alsoknown as alvocidib phosphate TP-1287 or alvocidib phosphate. TP-1287 isan orally bioavailable, highly soluble phosphate prodrug of alvocidib, apotent inhibitor of cyclin-dependent kinase-9 (CDK9), with potentialantineoplastic activity. Upon administration of the phosphate prodrugTP-1287, the prodrug is enzymatically cleaved at the tumor site and theactive moiety alvocidib is released. Alvocidib targets and binds toCDK9, thereby reducing the expression of CDK9 target genes such as theanti-apoptotic protein MCL-1, and inducing G1 cell cycle arrest andapoptosis in CDK9-overexpressing cancer cells. TP-1287 is disclosed,e.g., in Kim et al. Cancer Research (2017) Abstract 5133; Proceedings:AACR Annual Meeting 2017. In some embodiments, TP-1287 is administeredorally.

MDM2 Inhibitors

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with an MDM2 inhibitor. In someembodiments, the MDM2 inhibitor is chosen from idasanutlin, KRT-232,milademetan, or APG-115. In some embodiments, these combinations areused to treat the cancer indications disclosed herein, including thehematologic indications disclosed herein, including an MDS (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS). In someembodiments, these combinations are used to treat the cancer indicationsdisclosed herein, including the hematologic indications disclosedherein, including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary MDM2 Inhibitors

In some embodiments, the MDM2 inhibitor is a small molecule inhibitor.In some embodiments, the MDM2 inhibitor is idasanutlin. Idasanutlin isalso known as RG7388 or RO 5503781. Idasanutlin is an orally available,small molecule, antagonist of MDM2 (mouse double minute 2; Mdm2 p53binding protein homolog), with potential antineoplastic activity.Idasanutlin binds to MDM2 blocking the interaction between the MDM2protein and the transcriptional activation domain of the tumorsuppressor protein p53. By preventing the MDM2-p53 interaction, p53 isnot enzymatically degraded and the transcriptional activity of p53 isrestored, which may lead to p53-mediated induction of tumor cellapoptosis. Idasanutlin is disclosed, e.g., in Mascarenhas et al. Blood(2019) 134(6):525-533. In some embodiments, idasanutlin is administeredorally. In some embodiments, idasanutlin is administered on days 1-5 of,e.g., a 28 day cycle. In some embodiments, idasanutlin is administeredat 400-500 mg, e.g., 300 mg. In some embodiment, idasanutlin isadministered once or twice daily. In some embodiments, idasanutlin isadministered at 300 mg twice daily in cycle 1 (e.g., a 28 day cycle) oronce daily in cycles 2 and/or 3 (e.g., a 28 day cycle) for, e.g. 5 daysevery treatment cycle (e.g., a 28 day cycle).

In some embodiments, the MDM2 inhibitor is KRT-232. KRT-232 is alsoknown as(3R,5R,6S)-5-(3-Chlorophenyl)-6-(4-chlorophenyl)-3-methyl-1-((1S)-2-methyl-1-(((1-methylethyl)sulfonyl)methyl)propyl)-2-oxo-3-piperidineaceticAcid, or AMG-232. KRT-232 is an orally available inhibitor of MDM2(murine double minute 2), with potential antineoplastic activity. Uponoral administration, MDM2 inhibitor KRT-232 binds to the MDM2 proteinand prevents its binding to the transcriptional activation domain of thetumor suppressor protein p53. By preventing this MDM2-p53 interaction,the transcriptional activity of p53 is restored. KRT-232 is disclosed,e.g., in Garcia-Delgado et al. Blood (2019) 134(Supplement_1): 2945. Insome embodiments, KRT-232 is administered orally. In some embodiments,KRT-232 is administered once daily. In some embodiments, KRT-232 isadministered on days 1-7 of a cycle, e.g., a 28 day cycle. In someembodiments, KRT-232 is administered on days 4-10 and 18-24 of, e.g., a28 day cycle, for up to, e.g., 4 cycles.

In some embodiments, the MDM2 inhibitor is milademetan. Milademetan isalso known as HDM2 inhibitor DS-3032b or DS-3032b. Milademetan is anorally available MDM2 (murine double minute 2) antagonist with potentialantineoplastic activity. Upon oral administration, milademetan tosylatebinds to, and prevents the binding of MDM2 protein to thetranscriptional activation domain of the tumor suppressor protein p53.By preventing this MDM2-p53 interaction, the proteosome-mediatedenzymatic degradation of p53 is inhibited and the transcriptionalactivity of p53 is restored. This results in the restoration of p53signaling and leads to the p53-mediated induction of tumor cellapoptosis. Milademetan is disclosed, e.g., in DiNardo et al. Blood(2019) 134(Supplement_1):3932. In some embodiments, milademetan isadministered orally. In some embodiments, milademetan is administered at5-200 mg, e.g., 5 mg, 20 mg, 30 mg, 80 mg, 100 mg, 90 mg, and/or 200 mg.In some embodiments, milademetan is administered in a single capsule ormultiple capsules. In some embodiments, milademetan is administered at afixed dose. In some embodiments, milademetan is administered in a doseescalation regimen. In some embodiments, milademetan is administered infurther combination with quizartinib (an inhibitor of FLT3). In someembodiments, milademetan is administered at 5-200 mg (e.g., 5 mg, 20 mg,80 mg, or 200 mg), and quizartinib is administered at 20-30 mg (e.g., 20mg or 30 mg).

In some embodiments, the MDM2 inhibitor is APG-115. APG-115 is an orallyavailable inhibitor of human homologminute 2 (HDM2; mouse double minute2 homolog; MDM2), with potential antineoplastic activity. Upon oraladministration, the p53-HDM2 protein-protein interaction inhibitorAPG-115 binds to HDM2, preventing the binding of the HDM2 protein to thetranscriptional activation domain of the tumor suppressor protein p53.By preventing this HDM2-p53 interaction, the proteasome-mediatedenzymatic degradation of p53 is inhibited and the transcriptionalactivity of p53 is restored. This may result in the restoration of p53signaling and lead to the p53-mediated induction of tumor cellapoptosis. APG-115 is disclosed, e.g., in Fang et al. Journal forImmunoTherapy of Cancer (2019) 7(327). In some embodiments, APG-115 isadministered orally. In some embodiments, APG-115 is administered at100-250 mg, e.g., 100 mg, 150 mg, 200 mg, and/or 250 mg. In someembodiments, APG-115 is administered on days 1-5 of, e.g., a 28 daycycle. In some embodiments, APG-115 is administered on days 1-7 of,e.g., a 28 day cycle. In some embodiments, APG-115 is administered atflat dose. In some embodiments, APG-115 is administered on a doseescalation schedule. In some embodiments, APG-115 is administered at 100mg per day on day 1-5 of a 28 day cycle. In some embodiments, APG-115 isadministered at 150 mg per day on day 1-5 of a 28 day cycle. In someembodiments, APG-115 is administered at 200 mg per day on day 1-5 of a28 day cycle. In some embodiments, APG-115 is administered at 250 mg perday on day 1-5 of a 28 day cycle.

FLT3 Inhibitors

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with an FTL3 inhibitor. In someembodiments, the FLT3 inhibitor is chosen from gilteritinib,quizartinib, or crenolanib. In some embodiments, these combinations areused to treat the cancer indications disclosed herein, including thehematologic indications disclosed herein, including an MDS (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS). In someembodiments, these combinations are used to treat the cancer indicationsdisclosed herein, including the hematologic indications disclosedherein, including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary FLT3 Inhibitors

In some embodiments, the FLT3 inhibitor is gilteritinib. Gilteritinib isalso known as ASP2215. Gilteritinib is an orally bioavailable inhibitorof the receptor tyrosine kinases (RTKs) FMS-related tyrosine kinase 3(FLT3, STK1, or FLK2), AXL (UFO or JTK11) and anaplastic lymphoma kinase(ALK or CD246), with potential antineoplastic activity. Gilteritinibbinds to and inhibits both the wild-type and mutated forms of FLT3, AXLand ALK. This may result in an inhibition of FLT3, AXL, and ALK-mediatedsignal transduction pathways and reduction of tumor cell proliferationin cancer cell types that overexpress these RTKs. Gilteritinib isdisclosed, e.g, in Perl et al. N Engl J Med (2019) 381:1728-1740. Insome embodiments, gilteritinib is administered orally.

In some embodiments, the FLT3 inhibitor is quizartinib. Quizartinib isalso known as AC220 or1-(5-tert-butyl-1,2-oxazol-3-yl)-3-[4-[6-(2-morpholin-4-ylethoxy)imidazo[2,1-b][1,3]benzothiazol-2-yl]phenyl]urea.Quizartinib is disclosed, e.g., in Cortes et al. The Lancet (2019)20(7):984-997. In some embodiments, quizartinib is administered orally.In some embodiments, quizartinib is administered at 20-60 mg, e.g., 20mg, 30 mg, 40 mg, and/or 60 mg. In some embodiments, quizartinib isadministered once a day. In some embodiments, quizartinib isadministered at a flat dose. In some embodiments, quizartinib isadministered at 20 mg daily. In some embodiments, quizartinib isadministered at 30 mg once daily. In some embodiments, quizartinib isadministered at 40 mg once daily. In some embodiments, quizartinib isadministered in a dose escalation regimen. In some embodiments,quizartinib is administered at 30 mg daily for days 1-14 of, e.g., a 28day cycle, and is administered at 60 mg daily for days 15-28, of, e.g.,a 28 day cycle. In some embodiments, quizartinib is administered at 20mg daily for days 1-14 of, e.g., a 28 day cycle, and is administered at30 mg daily for days 15-28, of, e.g., a 28 day cycle.

In some embodiments, the FLT3 inhibitor is crenolanib. Crenolanib is anorally bioavailable small molecule, targeting the platelet-derivedgrowth factor receptor (PDGFR), with potential antineoplastic activity.Crenolanib binds to and inhibits PDGFR, which may result in theinhibition of PDGFR-related signal transduction pathways, and, so, theinhibition of tumor angiogenesis and tumor cell proliferation.Crenolanib is also known as CP-868596. Crenolanib is disclosed, e.g., inZimmerman et al. Blood (2013) 122(22):3607-3615. In some embodiments,crenolanib is administered orally. In some embodiments, crenolanib isadministered daily. In some embodiments, crenolanib is administered at100-200 mg, e.g., 100 mg or 200 mg. In some embodiments, crenolanib isadministered once a day, twice a day, or three times a day. In someembodiments, crenolanib is administered at 200 mg daily in three equaldoses, e.g., every 8 hours.

KIT Inhibitors

In certain embodiments, the anti-TIM-3 antibody described herein,optionally in combination with a hypomethylating agent described herein,is further administered in combination with a KIT inhibitor. In someembodiments, the KIT inhibitor is chosen from ripretinib, oravapritinib. In some embodiments, these combinations are used to treatthe cancer indications disclosed herein, including the hematologicindications disclosed herein, including an MDS (e.g., an intermediateMDS, a high risk MDS, or a very high risk MDS). In some embodiments,these combinations are used to treat the cancer indications disclosedherein, including the hematologic indications disclosed herein,including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary KIT Inhibitors

In some embodiments, the KIT inhibitor is ripretinib. Ripretinib is anorally bioavailable switch pocket control inhibitor of wild-type andmutated forms of the tumor-associated antigens (TAA) mast/stem cellfactor receptor (SCFR) KIT and platelet-derived growth factor receptoralpha (PDGFR-alpha; PDGFRa), with potential antineoplastic activity.Upon oral administration, ripretinib targets and binds to both wild-typeand mutant forms of KIT and PDGFRa specifically at their switch pocketbinding sites, thereby preventing the switch from inactive to activeconformations of these kinases and inactivating their wild-type andmutant forms. This abrogates KIT/PDGFRa-mediated tumor cell signalingand prevents proliferation in KIT/PDGFRa-driven cancers. DCC-2618 alsoinhibits several other kinases, including vascular endothelial growthfactor receptor type 2 (VEGFR2; KDR), angiopoietin-1 receptor (TIE2;TEK), PDGFR-beta and macrophage colony-stimulating factor 1 receptor(FMS; CSF1R), thereby further inhibiting tumor cell growth. Ripretinibis also known as DCC2618, QINLOCK™ (Deciphera), or1-N′-[2,5-difluoro-4-[2-(1-methylpyrazol-4-yl)pyridin-4-yl]oxyphenyl]-1-N′-phenylcyclopropane-1,1-dicarboxamide.In some embodiments, ripretinib is administered orally. In someembodiments, ripretinib is administered at 100-200 mg, e.g., 150 mg. Insome embodiments, ripretinib is administered in three 50 mg tablets. Insome embodiments, ripretinib is administered at 150 mg once daily. Insome embodiments, ripretinib is administered in three 50 mg tabletstaken together once daily.

In some embodiments, the KIT inhibitor is avapritinib. Avapritinib isalso known as BLU-285 or AYVAKIT™ (Blueprint Medicines). Avapritinib isan orally bioavailable inhibitor of specific mutated forms ofplatelet-derived growth factor receptor alpha (PDGFR alpha; PDGFRa) andmast/stem cell factor receptor c-Kit (SCFR), with potentialantineoplastic activity. Upon oral administration, avapritinibspecifically binds to and inhibits specific mutant forms of PDGFRa andc-Kit, including the PDGFRa D842V mutant and various KIT exon 17mutants. This results in the inhibition of PDGFRa- and c-Kit-mediatedsignal transduction pathways and the inhibition of proliferation intumor cells that express these PDGFRa and c-Kit mutants. In someembodiments, avapritinib is administered orally. In some embodiments,avapritinib is administered daily. In some embodiments, avapritinib isadministered at 100-300 mg, e.g., 100 mg, 200 mg, 300 mg. In someembodiments, avapritinib is administered once a day. In someembodiments, avapritinib is administered at 300 mg once a day. In someembodiments, avapritinib is administered at 200 mg once a day. In someembodiments, avapritinib is administered at 100 mg once a day. In someembodiments, avapritinib is administered continuously in, e.g., 28 daycycles.

PD-1 Inhibitors

In certain embodiments, the compounds and/or combinations describedherein are further administered in combination with a PD-1 inhibitor. Insome embodiments, the PD-1 inhibitor is chosen from spartalizumab(PDR001, Novartis), Nivolumab (Bristol-Myers Squibb), Pembrolizumab(Merck & Co), Pidilizumab (CureTech), MEDI0680 (Medimmune), REGN2810(Regeneron), TSR-042 (Tesaro), PF-06801591 (Pfizer), BGB-A317 (Beigene),BGB-108 (Beigene), INCSHR1210 (Incyte), or AMP-224 (Amplimmune). In someembodiments, these combinations are used to treat the cancer indicationsdisclosed herein, including the hematologic indications disclosedherein, including an MDS (e.g., an intermediate MDS, a high risk MDS, ora very high risk MDS). In some embodiments, these combinations are usedto treat the cancer indications disclosed herein, including thehematologic indications disclosed herein, including a CMML (e.g., aCMML-1 or a CMML-2).

Exemplary PD-1 Inhibitors

In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody molecule.In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody moleculeas described in US 2015/0210769, published on Jul. 30, 2015, entitled“Antibody Molecules to PD-1 and Uses Thereof,” incorporated by referencein its entirety. The antibody molecules described herein can be made byvectors, host cells, and methods described in US 2015/0210769,incorporated by reference in its entirety.

Other Exemplary PD-1 Inhibitors

In one embodiment, the anti-PD-1 antibody molecule is Nivolumab(Bristol-Myers Squibb), also known as MDX-1106, MDX-1106-04, ONO-4538,BMS-936558, or OPDIVO®. Nivolumab (clone 5C4) and other anti-PD-1antibodies are disclosed in U.S. Pat. No. 8,008,449 and WO 2006/121168,incorporated by reference in their entirety. In one embodiment, theanti-PD-1 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of Nivolumab.

In one embodiment, the anti-PD-1 antibody molecule is Pembrolizumab(Merck & Co), also known as Lambrolizumab, MK-3475, MK03475, SCH-900475,or KEYTRUDA®. Pembrolizumab and other anti-PD-1 antibodies are disclosedin Hamid, O. et al. (2013) New England Journal of Medicine 369 (2):134-44, U.S. Pat. No. 8,354,509, and WO 2009/114335, incorporated byreference in their entirety. In one embodiment, the anti-PD-1 antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of Pembrolizumab.

In one embodiment, the anti-PD-1 antibody molecule is Pidilizumab(CureTech), also known as CT-011. Pidilizumab and other anti-PD-1antibodies are disclosed in Rosenblatt, J. et al. (2011) J Immunotherapy34(5): 409-18, U.S. Pat. Nos. 7,695,715, 7,332,582, and 8,686,119,incorporated by reference in their entirety. In one embodiment, theanti-PD-1 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of Pidilizumab.

In one embodiment, the anti-PD-1 antibody molecule is MEDI0680(Medimmune), also known as AMP-514. MEDI0680 and other anti-PD-1antibodies are disclosed in U.S. Pat. No. 9,205,148 and WO 2012/145493,incorporated by reference in their entirety. In one embodiment, theanti-PD-1 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of MEDI0680.

In one embodiment, the anti-PD-1 antibody molecule is REGN2810(Regeneron). In one embodiment, the anti-PD-1 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of REGN2810.

In one embodiment, the anti-PD-1 antibody molecule is PF-06801591(Pfizer). In one embodiment, the anti-PD-1 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain or light chain variable region sequence, orthe heavy chain or light chain sequence of PF-06801591.

In one embodiment, the anti-PD-1 antibody molecule is BGB-A317 orBGB-108 (Beigene). In one embodiment, the anti-PD-1 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of BGB-A317 or BGB-108.

In one embodiment, the anti-PD-1 antibody molecule is INCSHR1210(Incyte), also known as INCSHR01210 or SHR-1210. In one embodiment, theanti-PD-1 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of INCSHR1210.

In one embodiment, the anti-PD-1 antibody molecule is TSR-042 (Tesaro),also known as ANB011. In one embodiment, the anti-PD-1 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of TSR-042.

Further known anti-PD-1 antibodies include those described, e.g., in WO2015/112800, WO 2016/092419, WO 2015/085847, WO 2014/179664, WO2014/194302, WO 2014/209804, WO 2015/200119, U.S. Pat. Nos. 8,735,553,7,488,802, 8,927,697, 8,993,731, and 9,102,727, incorporated byreference in their entirety.

In one embodiment, the anti-PD-1 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on PD-1 as, one ofthe anti-PD-1 antibodies described herein.

In one embodiment, the PD-1 inhibitor is a peptide that inhibits thePD-1 signaling pathway, e.g., as described in U.S. Pat. No. 8,907,053,incorporated by reference in its entirety. In one embodiment, the PD-1inhibitor is an immunoadhesin (e.g., an immunoadhesin comprising anextracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to aconstant region (e.g., an Fc region of an immunoglobulin sequence). Inone embodiment, the PD-1 inhibitor is AMP-224 (B7-DCIg (Amplimmune),e.g., disclosed in WO 2010/027827 and WO 2011/066342, incorporated byreference in their entirety).

PD-L1 Inhibitors

In certain embodiments, the compounds and/or combinations describedherein are further administered in combination with a PD-L1 inhibitor.In some embodiments, the PD-L1 inhibitor is chosen from FAZ053(Novartis), Atezolizumab (Genentech/Roche), Avelumab (Merck Serono andPfizer), Durvalumab (MedImmune/AstraZeneca), or BMS-936559(Bristol-Myers Squibb). In some embodiments, these combinations are usedto treat the cancer indications disclosed herein, including thehematologic indications disclosed herein, including an MDS (e.g., anintermediate MDS, a high risk MDS, or a very high risk MDS). In someembodiments, these combinations are used to treat the cancer indicationsdisclosed herein, including the hematologic indications disclosedherein, including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary PD-L1 Inhibitors

In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibodymolecule. In one embodiment, the PD-L1 inhibitor is an anti-PD-L1antibody molecule as disclosed in US 2016/0108123, published on Apr. 21,2016, entitled “Antibody Molecules to PD-L1 and Uses Thereof,”incorporated by reference in its entirety. The antibody moleculesdescribed herein can be made by vectors, host cells, and methodsdescribed in US 2016/0108123, incorporated by reference in its entirety.

Other Exemplary PD-L1 Inhibitors

In one embodiment, the anti-PD-L1 antibody molecule is Atezolizumab(Genentech/Roche), also known as MPDL3280A, RG7446, R05541267,YW243.55.570, or TECENTRIQ™. Atezolizumab and other anti-PD-L1antibodies are disclosed in U.S. Pat. No. 8,217,149, incorporated byreference in its entirety. In one embodiment, the anti-PD-L1 antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of Atezolizumab.

In one embodiment, the anti-PD-L1 antibody molecule is Avelumab (MerckSerono and Pfizer), also known as MSB0010718C. Avelumab and otheranti-PD-L1 antibodies are disclosed in WO 2013/079174, incorporated byreference in its entirety. In one embodiment, the anti-PD-L1 antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of Avelumab.

In one embodiment, the anti-PD-L1 antibody molecule is Durvalumab(MedImmune/AstraZeneca), also known as MEDI4736. Durvalumab and otheranti-PD-L1 antibodies are disclosed in U.S. Pat. No. 8,779,108,incorporated by reference in its entirety. In one embodiment, theanti-PD-L1 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of Durvalumab.

In one embodiment, the anti-PD-L1 antibody molecule is BMS-936559(Bristol-Myers Squibb), also known as MDX-1105 or 12A4. BMS-936559 andother anti-PD-L1 antibodies are disclosed in U.S. Pat. No. 7,943,743 andWO 2015/081158, incorporated by reference in their entirety. In oneembodiment, the anti-PD-L1 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain or light chain variable region sequence, or the heavy chain orlight chain sequence of BMS-936559.

Further known anti-PD-L1 antibodies include those described, e.g., in WO2015/181342, WO 2014/100079, WO 2016/000619, WO 2014/022758, WO2014/055897, WO 2015/061668, WO 2013/079174, WO 2012/145493, WO2015/112805, WO 2015/109124, WO 2015/195163, U.S. Pat. Nos. 8,168,179,8,552,154, 8,460,927, and 9,175,082, incorporated by reference in theirentirety.

In one embodiment, the anti-PD-L1 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on PD-L1 as, one ofthe anti-PD-L1 antibodies described herein.

LAG-3 Inhibitors

In certain embodiments, the compounds and/or combinations describedherein are further administered in combination with a LAG-3 inhibitor.In some embodiments, the LAG-3 inhibitor is chosen from LAG525(Novartis), BMS-986016 (Bristol-Myers Squibb), or TSR-033 (Tesaro). Insome embodiments, these combinations are used to treat the cancerindications disclosed herein, including the hematologic indicationsdisclosed herein, including an MDS (e.g., an intermediate MDS, a highrisk MDS, or a very high risk MDS). In some embodiments, thesecombinations are used to treat the cancer indications disclosed herein,including the hematologic indications disclosed herein, including a CMML(e.g., a CMML-1 or a CMML-2).

Exemplary LAG-3 Inhibitors

In one embodiment, the LAG-3 inhibitor is an anti-LAG-3 antibodymolecule. In one embodiment, the LAG-3 inhibitor is an anti-LAG-3antibody molecule as disclosed in US 2015/0259420, published on Sep. 17,2015, entitled “Antibody Molecules to LAG-3 and Uses Thereof,”incorporated by reference in its entirety. The antibody moleculesdescribed herein can be made by vectors, host cells, and methodsdescribed in US 2015/0259420, incorporated by reference in its entirety.

Other Exemplary LAG-3 Inhibitors

In one embodiment, the anti-LAG-3 antibody molecule is BMS-986016(Bristol-Myers Squibb), also known as BMS986016. BMS-986016 and otheranti-LAG-3 antibodies are disclosed in WO 2015/116539 and U.S. Pat. No.9,505,839, incorporated by reference in their entirety. In oneembodiment, the anti-LAG-3 antibody molecule comprises one or more ofthe CDR sequences (or collectively all of the CDR sequences), the heavychain or light chain variable region sequence, or the heavy chain orlight chain sequence of BMS-986016.

In one embodiment, the anti-LAG-3 antibody molecule is TSR-033 (Tesaro).In one embodiment, the anti-LAG-3 antibody molecule comprises one ormore of the CDR sequences (or collectively all of the CDR sequences),the heavy chain or light chain variable region sequence, or the heavychain or light chain sequence of TSR-033.

In one embodiment, the anti-LAG-3 antibody molecule is IMP731 orGSK2831781 (GSK and Prima BioMed). IMP731 and other anti-LAG-3antibodies are disclosed in WO 2008/132601 and U.S. Pat. No. 9,244,059,incorporated by reference in their entirety. In one embodiment, theanti-LAG-3 antibody molecule comprises one or more of the CDR sequences(or collectively all of the CDR sequences), the heavy chain or lightchain variable region sequence, or the heavy chain or light chainsequence of IMP731. In one embodiment, the anti-LAG-3 antibody moleculecomprises one or more of the CDR sequences (or collectively all of theCDR sequences), the heavy chain or light chain variable region sequence,or the heavy chain or light chain sequence of GSK2831781.

In one embodiment, the anti-LAG-3 antibody molecule is IMP761 (PrimaBioMed). In one embodiment, the anti-LAG-3 antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain or light chain variable region sequence, orthe heavy chain or light chain sequence of IMP761.

Further known anti-LAG-3 antibodies include those described, e.g., in WO2008/132601, WO 2010/019570, WO 2014/140180, WO 2015/116539, WO2015/200119, WO 2016/028672, U.S. Pat. Nos. 9,244,059, 9,505,839,incorporated by reference in their entirety.

In one embodiment, the anti-LAG-3 antibody is an antibody that competesfor binding with, and/or binds to the same epitope on LAG-3 as, one ofthe anti-LAG-3 antibodies described herein.

In one embodiment, the anti-LAG-3 inhibitor is a soluble LAG-3 protein,e.g., IMP321 (Prima BioMed), e.g., as disclosed in WO 2009/044273,incorporated by reference in its entirety.

GITR Agonists

In certain embodiments, the compounds and/or combinations describedherein are administered in combination with a GITR agonist. In someembodiments, the GITR agonist is GWN323 (NVS), BMS-986156, MK-4166 orMK-1248 (Merck), TRX518 (Leap Therapeutics), INCAGN1876 (Incyte/Agenus),AMG 228 (Amgen) or INBRX-110 (Inhibrx). In some embodiments, thesecombinations are used to treat the cancer indications disclosed herein,including the hematologic indications disclosed herein, including an MDS(e.g., an intermediate MDS, a high risk MDS, or a very high risk MDS).In some embodiments, these combinations are used to treat the cancerindications disclosed herein, including the hematologic indicationsdisclosed herein, including a CMML (e.g., a CMML-1 or a CMML-2).

Exemplary GITR Agonists

In one embodiment, the GITR agonist is an anti-GITR antibody molecule.In one embodiment, the GITR agonist is an anti-GITR antibody molecule asdescribed in WO 2016/057846, published on Apr. 14, 2016, entitled“Compositions and Methods of Use for Augmented Immune Response andCancer Therapy,” incorporated by reference in its entirety. The antibodymolecules described herein can be made by vectors, host cells, andmethods described in WO 2016/057846, incorporated by reference in itsentirety.

Other Exemplary GITR Agonists

In one embodiment, the anti-GITR antibody molecule is BMS-986156(Bristol-Myers Squibb), also known as BMS 986156 or BMS986156.BMS-986156 and other anti-GITR antibodies are disclosed, e.g., in U.S.Pat. No. 9,228,016 and WO 2016/196792, incorporated by reference intheir entirety.

In one embodiment, the anti-GITR antibody molecule comprises one or moreof the CDR sequences (or collectively all of the CDR sequences), theheavy chain or light chain variable region sequence, or the heavy chainor light chain sequence of BMS-986156.

In one embodiment, the anti-GITR antibody molecule is MK-4166 or MK-1248(Merck). MK-4166, MK-1248, and other anti-GITR antibodies are disclosed,e.g., in U.S. Pat. No. 8,709,424, WO 2011/028683, WO 2015/026684, andMahne et al. Cancer Res. 2017; 77(5):1108-1118, incorporated byreference in their entirety. In one embodiment, the anti-GITR antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of MK-4166 orMK-1248.

In one embodiment, the anti-GITR antibody molecule is TRX518 (LeapTherapeutics). TRX518 and other anti-GITR antibodies are disclosed,e.g., in U.S. Pat. Nos. 7,812,135, 8,388,967, 9,028,823, WO 2006/105021,and Ponte J et al. (2010) Clinical Immunology; 135:S96, incorporated byreference in their entirety. In one embodiment, the anti-GITR antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of TRX518.

In one embodiment, the anti-GITR antibody molecule is INCAGN1876(Incyte/Agenus). INCAGN1876 and other anti-GITR antibodies aredisclosed, e.g., in US 2015/0368349 and WO 2015/184099, incorporated byreference in their entirety. In one embodiment, the anti-GITR antibodymolecule comprises one or more of the CDR sequences (or collectively allof the CDR sequences), the heavy chain or light chain variable regionsequence, or the heavy chain or light chain sequence of INCAGN1876.

In one embodiment, the anti-GITR antibody molecule is AMG 228 (Amgen).AMG 228 and other anti-GITR antibodies are disclosed, e.g., in U.S. Pat.No. 9,464,139 and WO 2015/031667, incorporated by reference in theirentirety. In one embodiment, the anti-GITR antibody molecule comprisesone or more of the CDR sequences (or collectively all of the CDRsequences), the heavy chain or light chain variable region sequence, orthe heavy chain or light chain sequence of AMG 228.

In one embodiment, the anti-GITR antibody molecule is INBRX-110(Inhibrx). INBRX-110 and other anti-GITR antibodies are disclosed, e.g.,in US 2017/0022284 and WO 2017/015623, incorporated by reference intheir entirety. In one embodiment, the GITR agonist comprises one ormore of the CDR sequences (or collectively all of the CDR sequences),the heavy chain or light chain variable region sequence, or the heavychain or light chain sequence of INBRX-110.

In one embodiment, the GITR agonist (e.g., a fusion protein) is MEDI1873 (MedImmune), also known as MEDI1873. MEDI 1873 and other GITRagonists are disclosed, e.g., in US 2017/0073386, WO 2017/025610, andRoss et al. Cancer Res 2016; 76(14 Suppl): Abstract nr 561, incorporatedby reference in their entirety. In one embodiment, the GITR agonistcomprises one or more of an IgG Fc domain, a functional multimerizationdomain, and a receptor binding domain of a glucocorticoid-induced TNFreceptor ligand (GITRL) of MEDI 1873.

Further known GITR agonists (e.g., anti-GITR antibodies) include thosedescribed, e.g., in WO 2016/054638, incorporated by reference in itsentirety.

In one embodiment, the anti-GITR antibody is an antibody that competesfor binding with, and/or binds to the same epitope on GITR as, one ofthe anti-GITR antibodies described herein.

In one embodiment, the GITR agonist is a peptide that activates the GITRsignaling pathway. In one embodiment, the GITR agonist is animmunoadhesin binding fragment (e.g., an immunoadhesin binding fragmentcomprising an extracellular or GITR binding portion of GITRL) fused to aconstant region (e.g., an Fc region of an immunoglobulin sequence).

IL15/IL-15Ra Complexes

In certain embodiments, the compounds and/or combinations describedherein are further administered in combination with an IL-15/IL-15Racomplex. In some embodiments, the IL-15/IL-15Ra complex is chosen fromNIZ985 (Novartis), ATL-803 (Altor) or CYP0150 (Cytune). In someembodiments, these combinations are used to treat the cancer indicationsdisclosed herein, including the hematologic indications disclosedherein, including an MDS (e.g., an intermediate MDS, a high risk MDS, ora very high risk MDS). In some embodiments, these combinations are usedto treat the cancer indications disclosed herein, including thehematologic indications disclosed herein, including a CMML (e.g., aCMML-1 or a CMML-2).

Exemplary IL-15/IL-15Ra Complexes

In one embodiment, the IL-15/IL-15Ra complex comprises human IL-15complexed with a soluble form of human IL-15Ra. The complex may compriseIL-15 covalently or noncovalently bound to a soluble form of IL-15Ra. Ina particular embodiment, the human IL-15 is noncovalently bonded to asoluble form of IL-15Ra. In a particular embodiment, the human IL-15 ofthe composition comprises an amino acid sequence as described in WO2014/066527, incorporated herein by reference in its entirety, and thesoluble form of human IL-15Ra comprises an amino acid sequence, asdescribed in WO 2014/066527, incorporated by reference in its entirety.The molecules described herein can be made by vectors, host cells, andmethods described in WO 2007/084342, incorporated by reference in itsentirety.

Other Exemplary IL-15/IL-15Ra Complexes

In one embodiment, the IL-15/IL-15Ra complex is ALT-803, anIL-15/IL-15Ra Fc fusion protein (IL-15N72D:IL-15RaSu/Fc solublecomplex). ALT-803 is disclosed in WO 2008/143794, incorporated byreference in its entirety.

In one embodiment, the IL-15/IL-15Ra complex comprises IL-15 fused tothe sushi domain of IL-15Ra (CYP0150, Cytune). The sushi domain ofIL-15Ra refers to a domain beginning at the first cysteine residue afterthe signal peptide of IL-15Ra, and ending at the fourth cysteine residueafter said signal peptide. The complex of IL-15 fused to the sushidomain of IL-15Ra is disclosed in WO 2007/04606 and WO 2012/175222,incorporated by reference in their entirety.

Pharmaceutical Compositions, Formulations, and Kits

In another aspect, the disclosure provides compositions, e.g.,pharmaceutically acceptable compositions, which include a combinationdescribed herein, formulated together with a pharmaceutically acceptablecarrier. As used herein, “pharmaceutically acceptable carrier” includesany and all solvents, dispersion media, isotonic and absorption delayingagents, and the like that are physiologically compatible. The carriercan be suitable for intravenous, intramuscular, subcutaneous,parenteral, rectal, spinal or epidermal administration (e.g. byinjection or infusion).

The compositions described herein may be in a variety of forms. Theseinclude, for example, liquid, semi-solid and solid dosage forms, such asliquid solutions (e.g., injectable and infusible solutions), dispersionsor suspensions, liposomes and suppositories. The preferred form dependson the intended mode of administration and therapeutic application.Typical preferred compositions are in the form of injectable orinfusible solutions. The preferred mode of administration is parenteral(e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In apreferred embodiment, the antibody is administered by intravenousinfusion or injection. In another preferred embodiment, the antibody isadministered by intramuscular or subcutaneous injection.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal, epidural and intrasternal injection andinfusion.

Therapeutic compositions typically should be sterile and stable underthe conditions of manufacture and storage. The composition can beformulated as a solution, microemulsion, dispersion, liposome, or otherordered structure suitable to high antibody concentration. Sterileinjectable solutions can be prepared by incorporating the activecompound (e.g., antibody or antibody portion) in the required amount inan appropriate solvent with one or a combination of ingredientsenumerated above, as required, followed by filtered sterilization.Generally, dispersions are prepared by incorporating the active compoundinto a sterile vehicle that contains a basic dispersion medium and therequired other ingredients from those enumerated above. In the case ofsterile powders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum drying and freeze-dryingthat yields a powder of the active ingredient plus any additionaldesired ingredient from a previously sterile-filtered solution thereof.The proper fluidity of a solution can be maintained, for example, by theuse of a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.Prolonged absorption of injectable compositions can be brought about byincluding in the composition an agent that delays absorption, forexample, monostearate salts and gelatin.

A combination or a composition described herein can be formulated into aformulation (e.g., a dose formulation or dosage form) suitable foradministration (e.g., intravenous administration) to a subject asdescribed herein. The formulation described herein can be a liquidformulation, a lyophilized formulation, or a reconstituted formulation.

In certain embodiments, the formulation is a liquid formulation. In someembodiments, the formulation (e.g., liquid formulation) comprises aTIM-3 inhibitor (e.g., an anti-TIM-3 antibody molecule described herein)and a buffering agent.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of25 mg/mL to 250 mg/mL, e.g., 50 mg/mL to 200 mg/mL, 60 mg/mL to 180mg/mL, 70 mg/mL to 150 mg/mL, 80 mg/mL to 120 mg/mL, 90 mg/mL to 110mg/mL, 50 mg/mL to 150 mg/mL, 50 mg/mL to 100 mg/mL, 150 mg/mL to 200mg/mL, or 100 mg/mL to 200 mg/mL, e.g., 50 mg/mL, 60 mg/mL, 70 mg/mL, 80mg/mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL,or 150 mg/mL. In certain embodiments, the anti-TIM-3 antibody moleculeis present at a concentration of 80 mg/mL to 120 mg/mL, e.g., 100 mg/mL.

In some embodiments, the formulation (e.g., liquid formulation)comprises a buffering agent comprising histidine (e.g., a histidinebuffer). In certain embodiments, the buffering agent (e.g., histidinebuffer) is present at a concentration of 1 mM to 100 mM, e.g., 2 mM to50 mM, 5 mM to 40 mM, 10 mM to 30 mM, 15 to 25 mM, 5 mM to 40 mM, 5 mMto 30 mM, 5 mM to 20 mM, 5 mM to 10 mM, 40 mM to 50 mM, 30 mM to 50 mM,20 mM to 50 mM, 10 mM to 50 mM, or 5 mM to 50 mM, e.g., 2 mM, 5 mM, 10mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, or 50 mM. In someembodiments, the buffering agent (e.g., histidine buffer) is present ata concentration of 15 mM to 25 mM, e.g., 20 mM. In other embodiments,the buffering agent (e.g., a histidine buffer) has a pH of 4 to 7, e.g.,5 to 6, e.g., 5, 5.5, or 6. In some embodiments, the buffering agent(e.g., histidine buffer) has a pH of 5 to 6, e.g., 5.5. In certainembodiments, the buffering agent comprises a histidine buffer at aconcentration of 15 mM to 25 mM (e.g., 20 mM) and has a pH of 5 to 6(e.g., 5.5). In certain embodiments, the buffering agent compriseshistidine and histidine-HCl.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; and a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) andhas a pH of 5 to 6 (e.g., 5.5).

In some embodiments, the formulation (e.g., liquid formulation) furthercomprises a carbohydrate. In certain embodiments, the carbohydrate issucrose. In some embodiments, the carbohydrate (e.g., sucrose) ispresent at a concentration of 50 mM to 500 mM, e.g., 100 mM to 400 mM,150 mM to 300 mM, 180 mM to 250 mM, 200 mM to 240 mM, 210 mM to 230 mM,100 mM to 300 mM, 100 mM to 250 mM, 100 mM to 200 mM, 100 mM to 150 mM,300 mM to 400 mM, 200 mM to 400 mM, or 100 mM to 400 mM, e.g., 100 mM,150 mM, 180 mM, 200 mM, 220 mM, 250 mM, 300 mM, 350 mM, or 400 mM. Insome embodiments, the formulation comprises a carbohydrate or sucrosepresent at a concentration of 200 mM to 250 mM, e.g., 220 mM.

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) andhas a pH of 5 to 6 (e.g., 5.5); and a carbohydrate or sucrose present ata concentration of 200 mM to 250 mM, e.g., 220 mM.

In some embodiments, the formulation (e.g., liquid formulation) furthercomprises a surfactant. In certain embodiments, the surfactant ispolysorbate 20. In some embodiments, the surfactant or polysorbate 20)is present at a concentration of 0.005% to 0.1% (w/w), e.g., 0.01% to0.08%, 0.02% to 0.06%, 0.03% to 0.05%, 0.01% to 0.06%, 0.01% to 0.05%,0.01% to 0.03%, 0.06% to 0.08%, 0.04% to 0.08%, or 0.02% to 0.08% (w/w),e.g., 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or0.1% (w/w). In some embodiments, the formulation comprises a surfactantor polysorbate 20 present at a concentration of 0.03% to 0.05%, e.g.,0.04% (w/w).

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of80 to 120 mg/mL, e.g., 100 mg/mL; a buffering agent that comprises ahistidine buffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) andhas a pH of 5 to 6 (e.g., 5.5); a carbohydrate or sucrose present at aconcentration of 200 mM to 250 mM, e.g., 220 mM; and a surfactant orpolysorbate 20 present at a concentration of 0.03% to 0.05%, e.g., 0.04%(w/w).

In some embodiments, the formulation (e.g., liquid formulation)comprises an anti-TIM-3 antibody molecule present at a concentration of100 mg/mL; a buffering agent that comprises a histidine buffer (e.g.,histidine/histidine-HCL) at a concentration of 20 mM) and has a pH of5.5; a carbohydrate or sucrose present at a concentration of 220 mM; anda surfactant or polysorbate 20 present at a concentration of 0.04%(w/w).

In some embodiments, the liquid formulation is prepared by diluting aformulation comprising an anti-TIM-3 antibody molecule described herein.For example, a drug substance formulation can be diluted with a solutioncomprising one or more excipients (e.g., concentrated excipients). Insome embodiments, the solution comprises one, two, or all of histidine,sucrose, or polysorbate 20. In certain embodiments, the solutioncomprises the same excipient(s) as the drug substance formulation.Exemplary excipients include, but are not limited to, an amino acid(e.g., histidine), a carbohydrate (e.g., sucrose), or a surfactant(e.g., polysorbate 20). In certain embodiments, the liquid formulationis not a reconstituted lyophilized formulation. In other embodiments,the liquid formulation is a reconstituted lyophilized formulation. Insome embodiments, the formulation is stored as a liquid. In otherembodiments, the formulation is prepared as a liquid and then is dried,e.g., by lyophilization or spray-drying, prior to storage.

In certain embodiments, 0.5 mL to 10 mL (e.g., 0.5 mL to 8 mL, 1 mL to 6mL, or 2 mL to 5 mL, e.g., 1 mL, 1.2 mL, 1.5 mL, 2 mL, 3 mL, 4 mL, 4.5mL, or 5 mL) of the liquid formulation is filled per container (e.g.,vial). In other embodiments, the liquid formulation is filled into acontainer (e.g., vial) such that an extractable volume of at least 1 mL(e.g., at least 1.2 mL, at least 1.5 mL, at least 2 mL, at least 3 mL,at least 4 mL, or at least 5 mL) of the liquid formulation can bewithdrawn per container (e.g., vial). In certain embodiments, the liquidformulation is extracted from the container (e.g., vial) withoutdiluting at a clinical site. In certain embodiments, the liquidformulation is diluted from a drug substance formulation and extractedfrom the container (e.g., vial) at a clinical site. In certainembodiments, the formulation (e.g., liquid formulation) is injected toan infusion bag, e.g., within 1 hour (e.g., within 45 minutes, 30minutes, or 15 minutes) before the infusion starts to the patient.

A formulation described herein can be stored in a container. Thecontainer used for any of the formulations described herein can include,e.g., a vial, and optionally, a stopper, a cap, or both. In certainembodiments, the vial is a glass vial, e.g., a 6R white glass vial. Inother embodiments, the stopper is a rubber stopper, e.g., a grey rubberstopper. In other embodiments, the cap is a flip-off cap, e.g., analuminum flip-off cap. In some embodiments, the container comprises a 6Rwhite glass vial, a grey rubber stopper, and an aluminum flip-off cap.In some embodiments, the container (e.g., vial) is for a single-usecontainer. In certain embodiments, 25 mg/mL to 250 mg/mL, e.g., 50 mg/mLto 200 mg/mL, 60 mg/mL to 180 mg/mL, 70 mg/mL to 150 mg/mL, 80 mg/mL to120 mg/mL, 90 mg/mL to 110 mg/mL, 50 mg/mL to 150 mg/mL, 50 mg/mL to 100mg/mL, 150 mg/mL to 200 mg/mL, or 100 mg/mL to 200 mg/mL, e.g., 50mg/mL, 60 mg/mL, 70 mg/mL, 80 mg/mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120mg/mL, 130 mg/mL, 140 mg/mL, or 150 mg/mL, of the anti-TIM-3 antibodymolecule, is present in the container (e.g., vial).

In some embodiments, the formulation is a lyophilized formulation. Incertain embodiments, the lyophilized formulation is lyophilized or driedfrom a liquid formulation comprising an anti-TIM-3 antibody moleculedescribed herein. For example, 1 to 5 mL, e.g., 1 to 2 mL, of a liquidformulation can be filled per container (e.g., vial) and lyophilized.

In some embodiments, the formulation is a reconstituted formulation. Incertain embodiments, the reconstituted formulation is reconstituted froma lyophilized formulation comprising an anti-TIM-3 antibody moleculedescribed herein. For example, a reconstituted formulation can beprepared by dissolving a lyophilized formulation in a diluent such thatthe protein is dispersed in the reconstituted formulation. In someembodiments, the lyophilized formulation is reconstituted with 1 mL to 5mL, e.g., 1 mL to 2 mL, e.g., 1.2 mL, of water or buffer for injection.In certain embodiments, the lyophilized formulation is reconstitutedwith 1 mL to 2 mL of water for injection, e.g., at a clinical site.

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule (e.g., an anti-TIM-3 antibody moleculedescribed herein) and a buffering agent.

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule present at a concentration of 25 mg/mL to250 mg/mL, e.g., 50 mg/mL to 200 mg/mL, 60 mg/mL to 180 mg/mL, 70 mg/mLto 150 mg/mL, 80 mg/mL to 120 mg/mL, 90 mg/mL to 110 mg/mL, 50 mg/mL to150 mg/mL, 50 mg/mL to 100 mg/mL, 150 mg/mL to 200 mg/mL, or 100 mg/mLto 200 mg/mL, e.g., 50 mg/mL, 60 mg/mL, 70 mg/mL, 80 mg/mL, 90 mg/mL,100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL, or 150 mg/mL. Incertain embodiments, the anti-TIM-3 antibody molecule is present at aconcentration of 80 mg/mL to 120 mg/mL, e.g., 100 mg/mL.

In some embodiments, the reconstituted formulation comprises a bufferingagent comprising histidine (e.g., a histidine buffer). In certainembodiments, the buffering agent (e.g., histidine buffer) is present ata concentration of 1 mM to 100 mM, e.g., 2 mM to 50 mM, 5 mM to 40 mM,10 mM to 30 mM, 15 to 25 mM, 5 mM to 40 mM, 5 mM to 30 mM, 5 mM to 20mM, 5 mM to 10 mM, 40 mM to 50 mM, 30 mM to 50 mM, 20 mM to 50 mM, 10 mMto 50 mM, or 5 mM to 50 mM, e.g., 2 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25mM, 30 mM, 35 mM, 40 mM, 45 mM, or 50 mM. In some embodiments, thebuffering agent (e.g., histidine buffer) is present at a concentrationof 15 mM to 25 mM, e.g., 20 mM. In other embodiments, the bufferingagent (e.g., a histidine buffer) has a pH of 4 to 7, e.g., 5 to 6, e.g.,5, 5.5, or 6. In some embodiments, the buffering agent (e.g., histidinebuffer) has a pH of 5 to 6, e.g., 5.5. In certain embodiments, thebuffering agent comprises a histidine buffer at a concentration of 15 mMto 25 mM (e.g., 20 mM) and has a pH of 5 to 6 (e.g., 5.5). In certainembodiments, the buffering agent comprises histidine and histidine-HCl.

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule present at a concentration of 80 to 120mg/mL, e.g., 100 mg/mL; and a buffering agent that comprises a histidinebuffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) and has a pHof 5 to 6 (e.g., 5.5).

In some embodiments, the reconstituted formulation further comprises acarbohydrate. In certain embodiments, the carbohydrate is sucrose. Insome embodiments, the carbohydrate (e.g., sucrose) is present at aconcentration of 50 mM to 500 mM, e.g., 100 mM to 400 mM, 150 mM to 300mM, 180 mM to 250 mM, 200 mM to 240 mM, 210 mM to 230 mM, 100 mM to 300mM, 100 mM to 250 mM, 100 mM to 200 mM, 100 mM to 150 mM, 300 mM to 400mM, 200 mM to 400 mM, or 100 mM to 400 mM, e.g., 100 mM, 150 mM, 180 mM,200 mM, 220 mM, 250 mM, 300 mM, 350 mM, or 400 mM. In some embodiments,the formulation comprises a carbohydrate or sucrose present at aconcentration of 200 mM to 250 mM, e.g., 220 mM.

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule present at a concentration of 80 to 120mg/mL, e.g., 100 mg/mL; a buffering agent that comprises a histidinebuffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) and has a pHof 5 to 6 (e.g., 5.5); and a carbohydrate or sucrose present at aconcentration of 200 mM to 250 mM, e.g., 220 mM.

In some embodiments, the reconstituted formulation further comprises asurfactant. In certain embodiments, the surfactant is polysorbate 20. Insome embodiments, the surfactant or polysorbate 20) is present at aconcentration of 0.005% to 0.1% (w/w), e.g., 0.01% to 0.08%, 0.02% to0.06%, 0.03% to 0.05%, 0.01% to 0.06%, 0.01% to 0.05%, 0.01% to 0.03%,0.06% to 0.08%, 0.04% to 0.08%, or 0.02% to 0.08% (w/w), e.g., 0.01%,0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, or 0.1% (w/w).In some embodiments, the formulation comprises a surfactant orpolysorbate 20 present at a concentration of 0.03% to 0.05%, e.g., 0.04%(w/w).

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule present at a concentration of 80 to 120mg/mL, e.g., 100 mg/mL; a buffering agent that comprises a histidinebuffer at a concentration of 15 mM to 25 mM (e.g., 20 mM) and has a pHof 5 to 6 (e.g., 5.5); a carbohydrate or sucrose present at aconcentration of 200 mM to 250 mM, e.g., 220 mM; and a surfactant orpolysorbate 20 present at a concentration of 0.03% to 0.05%, e.g., 0.04%(w/w).

In some embodiments, the reconstituted formulation comprises ananti-TIM-3 antibody molecule present at a concentration of 100 mg/mL; abuffering agent that comprises a histidine buffer (e.g.,histidine/histidine-HCL) at a concentration of 20 mM) and has a pH of5.5; a carbohydrate or sucrose present at a concentration of 220 mM; anda surfactant or polysorbate 20 present at a concentration of 0.04%(w/w).

In some embodiments, the formulation is reconstituted such that anextractable volume of at least 1 mL (e.g., at least 1.2 mL, 1.5 mL, 2mL, 2.5 mL, or 3 mL) of the reconstituted formulation can be withdrawnfrom the container (e.g., vial) containing the reconstitutedformulation. In certain embodiments, the formulation is reconstitutedand/or extracted from the container (e.g., vial) at a clinical site. Incertain embodiments, the formulation (e.g., reconstituted formulation)is injected to an infusion bag, e.g., within 1 hour (e.g., within 45minutes, 30 minutes, or 15 minutes) before the infusion starts to thepatient.

Other exemplary buffering agents that can be used in the formulationdescribed herein include, but are not limited to, an arginine buffer, acitrate buffer, or a phosphate buffer. Other exemplary carbohydratesthat can be used in the formulation described herein include, but arenot limited to, trehalose, mannitol, sorbitol, or a combination thereof.The formulation described herein may also contain a tonicity agent,e.g., sodium chloride, and/or a stabilizing agent, e.g., an amino acid(e.g., glycine, arginine, methionine, or a combination thereof).

The antibody molecules can be administered by a variety of methods knownin the art, although for many therapeutic applications, the preferredroute/mode of administration is intravenous injection or infusion. Forexample, the antibody molecules can be administered by intravenousinfusion at a rate of more than 20 mg/min, e.g., 20-40 mg/min, andtypically greater than or equal to 40 mg/min to reach a dose of about 35to 440 mg/m², typically about 70 to 310 mg/m², and more typically, about110 to 130 mg/m². In embodiments, the antibody molecules can beadministered by intravenous infusion at a rate of less than 10 mg/min;preferably less than or equal to 5 mg/min to reach a dose of about 1 to100 mg/m², preferably about 5 to 50 mg/m², about 7 to 25 mg/m² and morepreferably, about 10 mg/m². As will be appreciated by the skilledartisan, the route and/or mode of administration will vary dependingupon the desired results. In certain embodiments, the active compoundmay be prepared with a carrier that will protect the compound againstrapid release, such as a controlled release formulation, includingimplants, transdermal patches, and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, and polylactic acid. Many methods for the preparationof such formulations are patented or generally known to those skilled inthe art. See, e.g., Sustained and Controlled Release Drug DeliverySystems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978.

In certain embodiments, an antibody molecule can be orally administered,for example, with an inert diluent or an assimilable edible carrier. Thecompound (and other ingredients, if desired) may also be enclosed in ahard or soft-shell gelatin capsule, compressed into tablets, orincorporated directly into the subject's diet. For oral therapeuticadministration, the compounds may be incorporated with excipients andused in the form of ingestible tablets, buccal tablets, troches,capsules, elixirs, suspensions, syrups, wafers, and the like. Toadminister a compound of the invention by other than parenteraladministration, it may be necessary to coat the compound with, orco-administer the compound with, a material to prevent its inactivation.Therapeutic compositions can also be administered with medical devicesknown in the art.

Dosage regimens are adjusted to provide the optimum desired response(e.g., a therapeutic response). For example, a single bolus may beadministered, several divided doses may be administered over time or thedose may be proportionally reduced or increased as indicated by theexigencies of the therapeutic situation. It is especially advantageousto formulate parenteral compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used hereinrefers to physically discrete units suited as unitary dosages for thesubjects to be treated; each unit contains a predetermined quantity ofactive compound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier. The specificationfor the dosage unit forms of the invention are dictated by and directlydependent on (a) the unique characteristics of the active compound andthe particular therapeutic effect to be achieved, and (b) thelimitations inherent in the art of compounding such an active compoundfor the treatment of sensitivity in individuals.

An exemplary, non-limiting range for a therapeutically orprophylactically effective amount of an antibody molecule is 50 mg to1500 mg, typically 100 mg to 1000 mg. In certain embodiments, theanti-TIM-3 antibody molecule is administered by injection (e.g.,subcutaneously or intravenously) at a dose (e.g., a flat dose) of about300 mg to about 500 mg (e.g., about 400 mg) or about 700 mg to about 900mg (e.g., about 800 mg). The dosing schedule (e.g., flat dosingschedule) can vary from e.g., once a week to once every 2, 3, 4, 5, or 6weeks. In one embodiment, the anti-TIM-3 antibody molecule isadministered at a dose from about 300 mg to 500 mg (e.g., about 400 mg)once every two weeks or once every four weeks. In one embodiment, theanti-TIM-3 antibody molecule is administered at a dose from about 700 mgto about 900 mg (e.g., about 800 mg) once every two weeks or once everyfour weeks. While not wishing to be bound by theory, in someembodiments, flat or fixed dosing can be beneficial to patients, forexample, to save drug supply and to reduce pharmacy errors.

The antibody molecule can be administered by intravenous infusion at arate of more than 20 mg/min, e.g., 20-40 mg/min, and typically greaterthan or equal to 40 mg/min to reach a dose of about 35 to 440 mg/m²,typically about 70 to 310 mg/m², and more typically, about 110 to 130mg/m². In embodiments, the infusion rate of about 110 to 130 mg/m²achieves a level of about 3 mg/kg. In other embodiments, the antibodymolecule can be administered by intravenous infusion at a rate of lessthan 10 mg/min, e.g., less than or equal to 5 mg/min to reach a dose ofabout 1 to 100 mg/m², e.g., about 5 to 50 mg/m², about 7 to 25 mg/m²,or, about 10 mg/m². In some embodiments, the antibody is infused over aperiod of about 30 min. It is to be noted that dosage values may varywith the type and severity of the condition to be alleviated. It is tobe further understood that for any particular subject, specific dosageregimens should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the compositions, and that dosage ranges set forthherein are exemplary only and are not intended to limit the scope orpractice of the claimed composition.

In some embodiments, the anti-TIM-3 antibody is administered incombination with a hypomethylating agent described herein. An exemplary,non-limiting range for a therapeutically or prophylactically effectiveamount of a hypomethylating agent is 50 mg/m² to about 100 mg/m²,typically 60 mg/m² to 80 mg/m². In certain embodiments, thehypomethylating agent is administered by injection (e.g., subcutaneouslyor intravenously) at a dose of about 50 mg/m² to about 60 mg/m² (about75 mg/m²), about 60 mg/m² to about 70 mg/m² (about 75 mg/m²), about 70mg/m² to about 80 mg/m² (about 85 mg/m²), about 80 mg/m² to about 90mg/m² (about 95 mg/m²), or about 90 mg/m² to about 100 mg/m² (about 95mg/m²). In some embodiments, the dosing schedule (e.g., flat dosingschedule) can vary during a 28-day cycle, from e.g., once a day for days1-7, or once a day for days 1-5, 8 and 9.

In one embodiment, azacitidine is administered intravenous orsubcutaneous at 75 mg/m² on Days 1-7 (or on Days 1 to 5 and Days 8 and9), and MBG453 is administered intravenously at 800 mg on Day 8 (Q4W) ofevery 28-day cycle.

The pharmaceutical compositions of the invention may include a“therapeutically effective amount” or a “prophylactically effectiveamount” of an antibody or antibody portion of the invention. A“therapeutically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredtherapeutic result. A therapeutically effective amount of the modifiedantibody or antibody fragment may vary according to factors such as thedisease state, age, sex, and weight of the individual, and the abilityof the antibody or antibody portion to elicit a desired response in theindividual. A therapeutically effective amount is also one in which anytoxic or detrimental effects of the modified antibody or antibodyfragment is outweighed by the therapeutically beneficial effects. A“therapeutically effective dosage” preferably inhibits a measurableparameter, e.g., tumor growth rate by at least about 20%, morepreferably by at least about 40%, even more preferably by at least about60%, and still more preferably by at least about 80% relative tountreated subjects. The ability of a compound to inhibit a measurableparameter, e.g., cancer, can be evaluated in an animal model systempredictive of efficacy in human tumors. Alternatively, this property ofa composition can be evaluated by examining the ability of the compoundto inhibit, such inhibition in vitro by assays known to the skilledpractitioner.

A “prophylactically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredprophylactic result. Typically, since a prophylactic dose is used insubjects prior to or at an earlier stage of disease, theprophylactically effective amount will be less than the therapeuticallyeffective amount.

Also within the scope of the disclosure is a kit comprising acombination, composition, or formulation described herein. The kit caninclude one or more other elements including: instructions for use(e.g., in accordance a dosage regimen described herein); other reagents,e.g., a label, a therapeutic agent, or an agent useful for chelating, orotherwise coupling, an antibody to a label or therapeutic agent, or aradioprotective composition; devices or other materials for preparingthe antibody for administration; pharmaceutically acceptable carriers;and devices or other materials for administration to a subject.

Use of the Combinations

The combinations described herein can be used to modify an immuneresponse in a subject. In some embodiments, the immune response isenhanced, stimulated or up-regulated. In certain embodiments, the immuneresponse is inhibited, reduced, or down-regulated. For example, thecombinations can be administered to cells in culture, e.g. in vitro orex vivo, or in a subject, e.g., in vivo, to treat, prevent, and/ordiagnose a variety of disorders, such as cancers and immune disorders.In some embodiments, the combination results in a synergistic effect. Inother embodiments, the combination results in an additive effect.

As used herein, the term “subject” is intended to include human andnon-human animals. In some embodiments, the subject is a human subject,e.g., a human patient having a disorder or condition characterized byabnormal TIM-3 functioning. Generally, the subject has at least someTIM-3 protein, including the TIM-3 epitope that is bound by the antibodymolecule, e.g., a high enough level of the protein and epitope tosupport antibody binding to TIM-3. The term “non-human animals” includesmammals and non-mammals, such as non-human primates. In someembodiments, the subject is a human. In some embodiments, the subject isa human patient in need of enhancement of an immune response. Thecombinations described herein are suitable for treating human patientshaving a disorder that can be treated by modulating (e.g., augmenting orinhibiting) an immune response. In certain embodiments, the patient hasor is at risk of having a disorder described herein, e.g., a cancerdescribed herein.

In some embodiments, the combination is used to treat a myelodysplasticsyndrome (MDS) (e.g., an intermediate MDS, a high risk MDS, or a veryhigh risk MDS), a chronic myelomonocytic leukemia (CMML) (e.g., CMML-1or CMML-2), a leukemia (e.g., an acute myeloid leukemia (AML), e.g., arelapsed or refractory AML or a de novo AML; or a chronic lymphocyticleukemia (CLL)), a lymphoma (e.g., T-cell lymphoma, B-cell lymphoma, anon-Hodgkin lymphoma, or a small lymphocytic lymphoma (SLL)), a myeloma(e.g., multiple myeloma), a lung cancer (e.g., a non-small cell lungcancer (NSCLC) (e.g., a NSCLC with squamous and/or non-squamoushistology, or a NSCLC adenocarcinoma), or a small cell lung cancer(SCLC)), a skin cancer (e.g., a Merkel cell carcinoma or a melanoma(e.g., an advanced melanoma)), an ovarian cancer, a mesothelioma, abladder cancer, a soft tissue sarcoma (e.g., a hemangiopericytoma(HPC)), a bone cancer (a bone sarcoma), a kidney cancer (e.g., a renalcancer (e.g., a renal cell carcinoma)), a liver cancer (e.g., ahepatocellular carcinoma), a cholangiocarcinoma, a sarcoma, amyelodysplastic syndrome (MDS), a prostate cancer, a breast cancer(e.g., a breast cancer that does not express one, two or all of estrogenreceptor, progesterone receptor, or Her2/neu, e.g., a triple negativebreast cancer), a colorectal cancer, a nasopharyngeal cancer, a duodenalcancer, an endometrial cancer, a pancreatic cancer, a head and neckcancer (e.g., head and neck squamous cell carcinoma (HNSCC), an analcancer, a gastro-esophageal cancer, a thyroid cancer (e.g., anaplasticthyroid carcinoma), a cervical cancer, or a neuroendocrine tumor (NET)(e.g., an atypical pulmonary carcinoid tumor).

In some embodiments, the cancer is a hematological cancer, e.g., amyelodysplastic syndrome (MDS) (e.g., an intermediate MDS, a high riskMDS, or a very high risk MDS), a chronic myelomonocytic leukemia (CMML)(e.g., CMML-1 or CMML-2), a leukemia, a lymphoma, or a myeloma. Forexample, an combination described herein can be used to treat cancersmalignancies, and related disorders, including, but not limited to,e.g., a myelodysplastic syndrome (MDS), e.g., an intermediate MDS, ahigh risk MDS, or a very high risk MDS, a chronic myelomonocyticleukemia (CMML), e.g., CMML-1 or CMML-2, an acute leukemia, e.g., B-cellacute lymphoid leukemia (BALL), T-cell acute lymphoid leukemia (TALL),acute myeloid leukemia (AML), acute lymphoid leukemia (ALL); a chronicleukemia, e.g., chronic myelogenous leukemia (CML), chronic lymphocyticleukemia (CLL); an additional hematologic cancer or hematologiccondition, e.g., B cell prolymphocytic leukemia, blastic plasmacytoiddendritic cell neoplasm, Burkitt's lymphoma, diffuse large B celllymphoma, Follicular lymphoma, Hairy cell leukemia, small cell- or alarge cell-follicular lymphoma, malignant lymphoproliferativeconditions, MALT lymphoma, mantle cell lymphoma, Marginal zone lymphoma,multiple myeloma, myelodysplasia and myelodysplastic syndrome,non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendriticcell neoplasm, Waldenström macroglobulinemia, myelofibrosis, amyloidlight chain amyloidosis, chronic neutrophilic leukemia, essentialthrombocythemia, chronic eosinophilic leukemia, chronic myelomonocyticleukemia, Richter Syndrome, mixed phenotype acute leukemia, acutebiphenotypic leukemia, and “preleukemia” which are a diverse collectionof hematological conditions united by ineffective production (ordysplasia) of myeloid blood cells, and the like.

In some embodiments, the combination is used to treat a myelodysplasticsyndrome (MDS) (e.g., an intermediate risk MDS, a high risk MDS, or avery high risk MDS). In some embodiments, the subject is classified as asubject with intermediate risk MDS, high risk MDS, or very high riskMDS. In some embodiments, a score of greater than 3 but less than orequal to 4.5 points on the International Prognostic Scoring System(IPSS-R) is classified as intermediate risk MDS. In some embodiments, ascore of greater than 4.5 but less than or equal to 6 points on theInternational Prognostic Scoring System (IPSS-R) is classified as highrisk MDS. In some embodiments, a score of greater 6 points on theInternational Prognostic Scoring System (IPSS-R) is classified as veryhigh risk MDS.

In some embodiments, the combination is used to treat a chronicmyelomonocytic leukemia (CMML) (e.g., CMML-1 or CMML-2). In someembodiments, the subject is classified as a subject with CMML-1 orCMML-2. In some embodiments, a subject with about 2% to about 4% blastsin the peripheral blood and/or about 5% to about 9% blasts in the bonemarrow is classified as a subject with CMML-1. In some embodiments, asubject with about 5% to about 19% blasts in the peripheral blood and/orabout 10% to about 19% blasts in the bone marrow is classified as asubject with CMML-2.

In some embodiments, the subject is not suitable for a standardtherapeutic regimen with established benefit in patients with a cancerdescribed herein. In some embodiments, the subject is unfit for achemotherapy or a hematopoietic stem cell transplant (HSCT).

In certain embodiments, the subject has been identified as having TIM-3expression in tumor infiltrating lymphocytes. In other embodiments, thesubject does not have detectable level of TIM-3 expression in tumorinfiltrating lymphocytes.

In some embodiments, the combination disclosed herein results inimproved remission duration and/or leukemic clearance in the subject(e.g., a patient in remission). For example, the subject can have alevel of minimal residual disease (MRD) below about 1%, typically below0.1%, after the treatment. Methods for determining minimal residualdisease, e.g., including Next-Generation Sequencing (NGS) and/orMultiparameter Flow Cytometry for acute myeloid leukemia, are described,e.g., in Schuurhuis et al. Blood. 2018; 131(12): 1275-1291; Ravandi etal., Blood Adv. 2018; 2(11): 1356-1366, DiNardo et al. Blood. 2019;133(1):7-17. MRD can be measured in a patient at baseline (i.e. beforetreatment), during treatment, end of treatment, and/or until diseaseprogression.

Methods of Treating Cancer

In one aspect, the disclosure relates to treatment of a subject in vivousing a combination described herein, or a composition or formulationcomprising a combination described herein, such that growth of canceroustumors is inhibited or reduced.

In certain embodiments, the combination comprises a TIM-3 inhibitor, anda hypomethylating agent. In some embodiments, the TIM-3 inhibitor,and/or the hypomethylating agent is administered or used in accordancewith a dosage regimen disclosed herein. In certain embodiments, thecombination is administered in an amount effective to treat a cancer ora symptom thereof.

The combinations, compositions, or formulations described herein can beused alone to inhibit the growth of cancerous tumors. Alternatively, thecombinations, compositions, or formulations described herein can be usedin combination with one or more of: a standard of care treatment forcancer, another antibody or antigen-binding fragment thereof, animmunomodulator (e.g., an activator of a costimulatory molecule or aninhibitor of an inhibitory molecule); a vaccine, e.g., a therapeuticcancer vaccine; or other forms of cellular immunotherapy, as describedherein.

Accordingly, in one embodiment, the disclosure provides a method ofinhibiting growth of tumor cells in a subject, comprising administeringto the subject a therapeutically effective amount of a combinationdescribed herein, e.g., in accordance with a dosage regimen describedherein. In an embodiment, the combination is administered in the form ofa composition or formulation described herein.

In one embodiment, the combination is suitable for the treatment ofcancer in vivo. To achieve antigen-specific enhancement of immunity, thecombination can be administered together with an antigen of interest.When a combination described herein is administered the combination canbe administered in either order or simultaneously.

In another aspect, a method of treating a subject, e.g., reducing orameliorating, a hyperproliferative condition or disorder (e.g., acancer), e.g., solid tumor, a hematological cancer, soft tissue tumor,or a metastatic lesion, in a subject is provided. The method includesadministering to the subject a combination described herein, or acomposition or formulation comprising a combination described herein, inaccordance with a dosage regimen disclosed herein.

As used herein, the term “cancer” is meant to include all types ofcancerous growths or oncogenic processes, metastatic tissues ormalignantly transformed cells, tissues, or organs, irrespective ofhistopathological type or stage of invasiveness. Examples of cancerousdisorders include, but are not limited to, hematological cancers, solidtumors, soft tissue tumors, and metastatic lesions.

In certain embodiments, the cancer is a hematological cancer. Examplesof hematological cancers include, but are not limited to,myelodysplastic syndrome (MDS) (e.g., an intermediate MDS, a high riskMDS, or a very high risk MDS), a chronic myelomonocytic leukemia (CMML)(e.g., CMML-1 or CMML-2), acute myeloid leukemia, chronic lymphocyticleukemia, small lymphocytic lymphoma, multiple myeloma, acutelymphocytic leukemia, non-Hodgkin's lymphoma, Hodgkin's lymphoma, mantlecell lymphoma, follicular lymphoma, Waldenstrom's macroglobulinemia,B-cell lymphoma and diffuse large B-cell lymphoma, precursorB-lymphoblastic leukemia/lymphoma, B-cell chronic lymphocyticleukemia/small lymphocytic lymphoma, B-cell prolymphocytic leukemia,lymphoplasmacytic lymphoma, splenic marginal zone B-cell lymphoma (withor without villous lymphocytes), hairy cell leukemia, plasma cellmyeloma/plasmacytoma, extranodal marginal zone B-cell lymphoma of theMALT type, nodal marginal zone B-cell lymphoma (with or withoutmonocytoid B cells), Burkitt's lymphoma, precursor T-lymphoblasticlymphoma/leukemia, T-cell prolymphocytic leukemia, T-cell granularlymphocytic leukemia, aggressive NK cell leukemia, adult T-celllymphoma/leukemia (HTLV 1-positive), nasal-type extranodal NK/T-celllymphoma, enteropathy-type T-cell lymphoma, hepatosplenic γ-δ T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, mycosisfungoides/Sezary syndrome, anaplastic large cell lymphoma (T/null cell,primary cutaneous type), anaplastic large cell lymphoma (T-/null-cell,primary systemic type), peripheral T-cell lymphoma not otherwisecharacterized, angioimmunoblastic T-cell lymphoma, polycythemia vera(PV), myelodysplastic syndrome (MDS), indolent Non-Hodgkin's Lymphoma(iNHL), and aggressive Non-Hodgkin's Lymphoma (aNHL).

In some embodiments, the hematological cancer is a myelodysplasticsyndrome (MDS) (e.g., an intermediate MDS, a high risk MDS, or a veryhigh risk MDS), a chronic myelomonocytic leukemia (CMML) (e.g., CMML-1or CMML-2).

Examples of solid tumors include, but are not limited to, malignancies,e.g., sarcomas, and carcinomas (including adenocarcinomas and squamouscell carcinomas), of the various organ systems, such as those affectingliver, lung, breast, lymphoid, gastrointestinal (e.g., colon), anal,genitals and genitourinary tract (e.g., renal, urothelial, bladder),prostate, CNS (e.g., brain, neural or glial cells), head and neck, skin,pancreas, and pharynx. Adenocarcinomas include malignancies such as mostcolon cancers, rectal cancer, renal cancer (e.g., renal-cell carcinoma(e.g., clear cell or non-clear cell renal cell carcinoma), liver cancer,lung cancer (e.g., non-small cell carcinoma of the lung (e.g., squamousor non-squamous non-small cell lung cancer)), cancer of the smallintestine, and cancer of the esophagus. Squamous cell carcinomas includemalignancies, e.g., in the lung, esophagus, skin, head and neck region,oral cavity, anus, and cervix. In one embodiment, the cancer is amelanoma, e.g., an advanced stage melanoma. The cancer may be at anearly, intermediate, late stage or metastatic cancer. Metastatic lesionsof the aforementioned cancers can also be treated or prevented using thecombinations described herein.

In certain embodiments, the cancer is a solid tumor. In someembodiments, the cancer is an ovarian cancer. In other embodiments, thecancer is a lung cancer, e.g., a small cell lung cancer (SCLC) or anon-small cell lung cancer (NSCLC). In other embodiments, the cancer isa mesothelioma. In other embodiments, the cancer is a skin cancer, e.g.,a Merkel cell carcinoma or a melanoma. In other embodiments, the canceris a kidney cancer, e.g., a renal cell carcinoma (RCC). In otherembodiments, the cancer is a bladder cancer. In other embodiments, thecancer is a soft tissue sarcoma, e.g., a hemangiopericytoma (HPC). Inother embodiments, the cancer is a bone cancer, e.g., a bone sarcoma. Inother embodiments, the cancer is a colorectal cancer. In otherembodiments, the cancer is a pancreatic cancer. In other embodiments,the cancer is a nasopharyngeal cancer. In other embodiments, the canceris a breast cancer. In other embodiments, the cancer is a duodenalcancer. In other embodiments, the cancer is an endometrial cancer. Inother embodiments, the cancer is an adenocarcinoma, e.g., an unknownadenocarcinoma. In other embodiments, the cancer is a liver cancer,e.g., a hepatocellular carcinoma. In other embodiments, the cancer is acholangiocarcinoma. In other embodiments, the cancer is a sarcoma. Incertain embodiments, the cancer is a myelodysplastic syndrome (MDS)(e.g., a high risk MDS).

In another embodiment, the cancer is a carcinoma (e.g., advanced ormetastatic carcinoma), melanoma or a lung carcinoma, e.g., a non-smallcell lung carcinoma. In one embodiment, the cancer is a lung cancer,e.g., a non-small cell lung cancer or small cell lung cancer. In someembodiments, the non-small cell lung cancer is a stage I (e.g., stage Iaor Ib), stage II (e.g., stage IIa or IIb), stage III (e.g., stage Ma orMb), or stage IV, non-small cell lung cancer. In one embodiment, thecancer is a melanoma, e.g., an advanced melanoma. In one embodiment, thecancer is an advanced or unresectable melanoma that does not respond toother therapies. In other embodiments, the cancer is a melanoma with aBRAF mutation (e.g., a BRAF V600 mutation). In another embodiment, thecancer is a hepatocarcinoma, e.g., an advanced hepatocarcinoma, with orwithout a viral infection, e.g., a chronic viral hepatitis. In anotherembodiment, the cancer is a prostate cancer, e.g., an advanced prostatecancer. In yet another embodiment, the cancer is a myeloma, e.g.,multiple myeloma. In yet another embodiment, the cancer is a renalcancer, e.g., a renal cell carcinoma (RCC) (e.g., a metastatic RCC, anon-clear cell renal cell carcinoma (nccRCC), or clear cell renal cellcarcinoma (CCRCC)).

In some embodiments, the cancer is an MSI-high cancer. In someembodiments, the cancer is a metastatic cancer. In other embodiments,the cancer is an advanced cancer. In other embodiments, the cancer is arelapsed or refractory cancer.

Exemplary cancers whose growth can be inhibited using the combinations,compositions, or formulations, as disclosed herein, include cancerstypically responsive to immunotherapy. Additionally, refractory orrecurrent malignancies can be treated using the combinations describedherein.

Examples of other cancers that can be treated include, but are notlimited to, basal cell carcinoma, biliary tract cancer; bladder cancer;bone cancer; brain and CNS cancer; primary CNS lymphoma; neoplasm of thecentral nervous system (CNS); breast cancer; cervical cancer;choriocarcinoma; colon and rectum cancer; connective tissue cancer;cancer of the digestive system; endometrial cancer; esophageal cancer;eye cancer; cancer of the head and neck; gastric cancer;intra-epithelial neoplasm; kidney cancer; larynx cancer; leukemia(including acute myeloid leukemia, chronic myeloid leukemia, acutelymphoblastic leukemia, chronic lymphocytic leukemia, chronic or acuteleukemia); liver cancer; lung cancer (e.g., small cell and non-smallcell); lymphoma including Hodgkin's and non-Hodgkin's lymphoma;lymphocytic lymphoma; melanoma, e.g., cutaneous or intraocular malignantmelanoma; myeloma; neuroblastoma; oral cavity cancer (e.g., lip, tongue,mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer;retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of therespiratory system; sarcoma; skin cancer; stomach cancer; testicularcancer; thyroid cancer; uterine cancer; cancer of the urinary system,hepatocarcinoma, cancer of the anal region, carcinoma of the fallopiantubes, carcinoma of the vagina, carcinoma of the vulva, cancer of thesmall intestine, cancer of the endocrine system, cancer of theparathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue,cancer of the urethra, cancer of the penis, solid tumors of childhood,spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi'ssarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma,environmentally induced cancers including those induced by asbestos, aswell as other carcinomas and sarcomas, and combinations of said cancers.

As used herein, the term “subject” is intended to include human andnon-human animals. In some embodiments, the subject is a human subject,e.g., a human patient having a disorder or condition characterized byabnormal TIM-3 functioning. Generally, the subject has at least someTIM-3 protein, including the TIM-3 epitope that is bound by the antibodymolecule, e.g., a high enough level of the protein and epitope tosupport antibody binding to TIM-3. The term “non-human animals” includesmammals and non-mammals, such as non-human primates. In someembodiments, the subject is a human. In some embodiments, the subject isa human patient in need of enhancement of an immune response. Themethods and compositions described herein are suitable for treatinghuman patients having a disorder that can be treated by modulating(e.g., augmenting or inhibiting) an immune response.

Methods and compositions disclosed herein are useful for treatingmetastatic lesions associated with the aforementioned cancers.

In some embodiments, the method further comprises determining whether atumor sample is positive for one or more of PD-L1, CD8, and IFN-γ, andif the tumor sample is positive for one or more, e.g., two, or allthree, of the markers, then administering to the patient atherapeutically effective amount of an anti-TIM-3 antibody molecule,optionally in combination with one or more other immunomodulators oranti-cancer agents, as described herein.

In some embodiments, the combination described herein is used to treat acancer that expresses TIM-3. TIM-3-expressing cancers include, but arenot limited to, cervical cancer (Cao et al., PLoS One. 2013; 8(1):e53834), lung cancer (Zhuang et al., Am J Clin Pathol. 2012;137(6):978-985) (e.g., non-small cell lung cancer), acute myeloidleukemia (Kikushige et al., Cell Stem Cell. 2010 Dec. 3; 7(6):708-17),diffuse large B cell lymphoma, melanoma (Fourcade et al., JEM, 2010; 207(10): 2175), renal cancer (e.g., renal cell carcinoma (RCC), e.g.,kidney clear cell carcinoma, kidney papillary cell carcinoma, ormetastatic renal cell carcinoma), squamous cell carcinoma, esophagealsquamous cell carcinoma, nasopharyngeal carcinoma, colorectal cancer,breast cancer (e.g., a breast cancer that does not express one, two orall of estrogen receptor, progesterone receptor, or Her2/neu, e.g., atriple negative breast cancer), mesothelioma, hepatocellular carcinoma,and ovarian cancer. The TIM-3-expressing cancer may be a metastaticcancer.

In other embodiments, the combination described herein is used to treata cancer that is characterized by macrophage activity or high expressionof macrophage cell markers. In an embodiment, the combination is used totreat a cancer that is characterized by high expression of one or moreof the following macrophage cell markers: LILRB4 (macrophage inhibitoryreceptor), CD14, CD16, CD68, MSR1, SIGLEC1, TREM2, CD163, ITGAX, ITGAM,CD11b, or CD11c. Examples of such cancers include, but are not limitedto, diffuse large B-cell lymphoma, glioblastoma multiforme, kidney renalclear cell carcinoma, pancreatic adenocarcinoma, sarcoma, liverhepatocellular carcinoma, lung adenocarcinoma, kidney renal papillarycell carcinoma, skin cutaneous melanoma, brain lower grade glioma, lungsquamous cell carcinoma, ovarian serious cystadenocarcinoma, head andneck squamous cell carcinoma, breast invasive carcinoma, acute myeloidleukemia, cervical squamous cell carcinoma, endocervical adenocarcinoma,uterine carcinoma, colorectal cancer, uterine corpus endometrialcarcinoma, thyroid carcinoma, bladder urothelial carcinoma,adrenocortical carcinoma, kidney chromophobe, and prostateadenocarcinoma.

The combination therapies described herein can include a compositionco-formulated with, and/or co-administered with, one or more therapeuticagents, e.g., one or more anti-cancer agents, cytotoxic or cytostaticagents, hormone treatment, vaccines, and/or other immunotherapies. Inother embodiments, the antibody molecules are administered incombination with other therapeutic treatment modalities, includingsurgery, radiation, cryosurgery, and/or thermotherapy. Such combinationtherapies may advantageously utilize lower dosages of the administeredtherapeutic agents, thus avoiding possible toxicities or complicationsassociated with the various monotherapies.

The combinations, compositions, and formulations described herein can beused further in combination with other agents or therapeutic modalities,e.g., a second therapeutic agent chosen from one or more of the agentslisted in Table 6 of WO 2017/019897, the content of which isincorporated by reference in its entirety. In one embodiment, themethods described herein include administering to the subject ananti-TIM-3 antibody molecule as described in WO2017/019897 (optionallyin combination with one or more inhibitors of PD-1, PD-L1, LAG-3, CEACAM(e.g., CEACAM-1 and/or CEACAM-5), or CTLA-4)), further includeadministration of a second therapeutic agent chosen from one or more ofthe agents listed in Table 6 of WO 2017/019897, in an amount effectiveto treat or prevent a disorder, e.g., a disorder as described herein,e.g., a cancer. When administered in combination, the TIM-3 inhibitor,hypomethylating agent, one or more additional agents, or all, can beadministered in an amount or dose that is higher, lower or the same thanthe amount or dosage of each agent used individually, e.g., as amonotherapy. In certain embodiments, the administered amount or dosageof the TIM-3 inhibitor, hypomethylating agent, one or more additionalagents, or all, is lower (e.g., at least 20%, at least 30%, at least40%, or at least 50%) than the amount or dosage of each agent usedindividually, e.g., as a monotherapy. In other embodiments, the amountor dosage of the TIM-3 inhibitor, hypomethylating agent, one or moreadditional agents, or all, that results in a desired effect (e.g.,treatment of cancer) is lower (e.g., at least 20%, at least 30%, atleast 40%, or at least 50% lower).

In other embodiments, the additional therapeutic agent is chosen fromone or more of the agents listed in Table 6 of WO 2017/019897. In someembodiments, the additional therapeutic agent is chosen from one or moreof: 1) a protein kinase C (PKC) inhibitor; 2) a heat shock protein 90(HSP90) inhibitor; 3) an inhibitor of a phosphoinositide 3-kinase (PI3K)and/or target of rapamycin (mTOR); 4) an inhibitor of cytochrome P450(e.g., a CYP17 inhibitor or a 17alpha-Hydroxylase/C17-20 Lyaseinhibitor); 5) an iron chelating agent; 6) an aromatase inhibitor; 7) aninhibitor of p53, e.g., an inhibitor of a p53/Mdm2 interaction; 8) anapoptosis inducer; 9) an angiogenesis inhibitor; 10) an aldosteronesynthase inhibitor; 11) a smoothened (SMO) receptor inhibitor; 12) aprolactin receptor (PRLR) inhibitor; 13) a Wnt signaling inhibitor; 14)a CDK4/6 inhibitor; 15) a fibroblast growth factor receptor 2(FGFR2)/fibroblast growth factor receptor 4 (FGFR4) inhibitor; 16) aninhibitor of macrophage colony-stimulating factor (M-CSF); 17) aninhibitor of one or more of c-KIT, histamine release, Flt3 (e.g.,FLK2/STK1) or PKC; 18) an inhibitor of one or more of VEGFR-2 (e.g.,FLK-1/KDR), PDGFRbeta, c-KIT or Raf kinase C; 19) a somatostatin agonistand/or a growth hormone release inhibitor; 20) an anaplastic lymphomakinase (ALK) inhibitor; 21) an insulin-like growth factor 1 receptor(IGF-1R) inhibitor; 22) a P-Glycoprotein 1 inhibitor; 23) a vascularendothelial growth factor receptor (VEGFR) inhibitor; 24) a BCR-ABLkinase inhibitor; 25) an FGFR inhibitor; 26) an inhibitor of CYP11B2;27) a HDM2 inhibitor, e.g., an inhibitor of the HDM2-p53 interaction;28) an inhibitor of a tyrosine kinase; 29) an inhibitor of c-MET; 30) aninhibitor of JAK; 31) an inhibitor of DAC; 32) an inhibitor of11(3-hydroxylase; 33) an inhibitor of IAP; 34) an inhibitor of PIMkinase; 35) an inhibitor of Porcupine; 36) an inhibitor of BRAF, e.g.,BRAF V600E or wild-type BRAF; 37) an inhibitor of HER3; 38) an inhibitorof MEK; or 39) an inhibitor of a lipid kinase, e.g., as described inTable 6 of WO 2017/019897.

Additional embodiments of combination therapies comprising an anti-TIM-3antibody molecule described herein are described in WO2017/019897, whichis incorporated by reference in its entirety.

Nucleic Acids

In some embodiments, the combination described herein comprises ananti-TIM-3 antibody. The anti-TIM-3 antibody molecules described hereincan be encoded by nucleic acids described herein. The nucleic acids canbe used to produce the anti-TIM-3 antibody molecules described herein.

In certain embodiments, the nucleic acid comprises nucleotide sequencesthat encode heavy and light chain variable regions and CDRs of theanti-TIM-3 antibody molecules, as described herein. For example, thepresent disclosure features a first and second nucleic acid encodingheavy and light chain variable regions, respectively, of an anti-TIM-3antibody molecule chosen from one or more of the antibody moleculesdisclosed herein, e.g., an antibody of Tables 1-4 of US 2015/0218274.The nucleic acid can comprise a nucleotide sequence encoding any one ofthe amino acid sequences in the tables herein, or a sequencesubstantially identical thereto (e.g., a sequence at least about 85%,90%, 95%, 99% or more identical thereto, or which differs by no morethan 3, 6, 15, 30, or 45 nucleotides from the sequences provided inTables 1-4. For example, disclosed herein is a first and second nucleicacid encoding heavy and light chain variable regions, respectively, ofan anti-TIM-3 antibody molecule chosen from one or more of, e.g., any ofABTIM3, ABTIM3-hum01, ABTIM3-hum02, ABTIM3-hum03, ABTIM3-hum04,ABTIM3-hum05, ABTIM3-hum06, ABTIM3-hum07, ABTIM3-hum08, ABTIM3-hum09,ABTIM3-hum10, ABTIM3-hum11, ABTIM3-hum12, ABTIM3-hum13, ABTIM3-hum14,ABTIM3-hum15, ABTIM3-hum16, ABTIM3-hum17, ABTIM3-hum18, ABTIM3-hum19,ABTIM3-hum20, ABTIM3-hum21, ABTIM3-hum22, ABTIM3-hum23, as summarized inTables 1-4, or a sequence substantially identical thereto.

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs from a heavy chainvariable region having an amino acid sequence as set forth in Tables1-4, or a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or havingone or more substitutions, e.g., conserved substitutions). In someembodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, or three CDRs from a light chain variableregion having an amino acid sequence as set forth in Tables 1-4, or asequence substantially homologous thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, and/or having one ormore substitutions, e.g., conserved substitutions). In some embodiments,the nucleic acid can comprise a nucleotide sequence encoding at leastone, two, three, four, five, or six CDRs from heavy and light chainvariable regions having an amino acid sequence as set forth in Tables1-4, or a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or havingone or more substitutions, e.g., conserved substitutions).

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs from a heavy chainvariable region having the nucleotide sequence as set forth in Tables1-4, a sequence substantially homologous thereto (e.g., a sequence atleast about 85%, 90%, 95%, 99% or more identical thereto, and/or capableof hybridizing under the stringency conditions described herein). Insome embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, or three CDRs from a light chain variableregion having the nucleotide sequence as set forth in Tables 1-4, or asequence substantially homologous thereto (e.g., a sequence at leastabout 85%, 90%, 95%, 99% or more identical thereto, and/or capable ofhybridizing under the stringency conditions described herein). Incertain embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, three, four, five, or six CDRs from heavyand light chain variable regions having the nucleotide sequence as setforth in Tables 1-4, or a sequence substantially homologous thereto(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, and/or capable of hybridizing under the stringency conditionsdescribed herein). The nucleic acids disclosed herein includedeoxyribonucleotides or ribonucleotides, or analogs thereof. Thepolynucleotide may be either single-stranded or double-stranded, and ifsingle-stranded may be the coding strand or non-coding (antisense)strand. A polynucleotide may comprise modified nucleotides, such asmethylated nucleotides and nucleotide analogs. The sequence ofnucleotides may be interrupted by non-nucleotide components. Apolynucleotide may be further modified after polymerization, such as byconjugation with a labeling component. The nucleic acid may be arecombinant polynucleotide, or a polynucleotide of genomic, cDNA,semisynthetic, or synthetic origin which either does not occur in natureor is linked to another polynucleotide in a nonnatural arrangement.

In certain embodiments, the nucleotide sequence that encodes theanti-TIM-3 antibody molecule is codon optimized.

In some embodiments, nucleic acids comprising nucleotide sequences thatencode heavy and light chain variable regions and CDRs of the anti-TIM-3antibody molecules, as described herein, are disclosed. For example, thedisclosure provides a first and second nucleic acid encoding heavy andlight chain variable regions, respectively, of an anti-TIM-3 antibodymolecule according to Tables 1-4 or a sequence substantially identicalthereto. For example, the nucleic acid can comprise a nucleotidesequence encoding an anti-TIM-3 antibody molecule according to Table1-4, or a sequence substantially identical to that nucleotide sequence(e.g., a sequence at least about 85%, 90%, 95%, 99% or more identicalthereto, or which differs by no more than 3, 6, 15, 30, or 45nucleotides from the aforementioned nucleotide sequence.

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs, or hypervariableloops, from a heavy chain variable region having an amino acid sequenceas set forth in Tables 1-4, or a sequence substantially homologousthereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or moreidentical thereto, and/or having one, two, three or more substitutions,insertions or deletions, e.g., conserved substitutions).

In certain embodiments, the nucleic acid can comprise a nucleotidesequence encoding at least one, two, or three CDRs, or hypervariableloops, from a light chain variable region having an amino acid sequenceas set forth in Tables 1-4, or a sequence substantially homologousthereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or moreidentical thereto, and/or having one, two, three or more substitutions,insertions or deletions, e.g., conserved substitutions).

In some embodiments, the nucleic acid can comprise a nucleotide sequenceencoding at least one, two, three, four, five, or six CDRs, orhypervariable loops, from heavy and light chain variable regions havingan amino acid sequence as set forth in Table 1-4, or a sequencesubstantially homologous thereto (e.g., a sequence at least about 85%,90%, 95%, 99% or more identical thereto, and/or having one, two, threeor more substitutions, insertions or deletions, e.g., conservedsubstitutions).

In some embodiments, the anti-TIM-3 antibody molecule is isolated orrecombinant.

In some aspects, the application features host cells and vectorscontaining the nucleic acids described herein. The nucleic acids may bepresent in a single vector or separate vectors present in the same hostcell or separate host cell, as described in more detail herein.

Vectors and Host Cells

In some embodiments, the combination described herein comprises ananti-TIM-3 antibody molecule. The anti-TIM-3 antibody moleculesdescribed herein can be produced using host cells and vectors containingthe nucleic acids described herein. The nucleic acids may be present ina single vector or separate vectors present in the same host cell orseparate host cell.

In one embodiment, the vectors comprise nucleotides encoding an antibodymolecule described herein. In one embodiment, the vectors comprise thenucleotide sequences described herein. The vectors include, but are notlimited to, a virus, plasmid, cosmid, lambda phage or a yeast artificialchromosome (YAC).

Numerous vector systems can be employed. For example, one class ofvectors utilizes DNA elements which are derived from animal viruses suchas, for example, bovine papilloma virus, polyoma virus, adenovirus,vaccinia virus, baculovirus, retroviruses (Rous Sarcoma Virus, MMTV orMOMLV) or SV40 virus. Another class of vectors utilizes RNA elementsderived from RNA viruses such as Semliki Forest virus, Eastern EquineEncephalitis virus and Flaviviruses.

Additionally, cells which have stably integrated the DNA into theirchromosomes may be selected by introducing one or more markers whichallow for the selection of transfected host cells. The marker mayprovide, for example, prototropy to an auxotrophic host, biocideresistance (e.g., antibiotics), or resistance to heavy metals such ascopper, or the like. The selectable marker gene can be either directlylinked to the DNA sequences to be expressed or introduced into the samecell by cotransformation. Additional elements may also be needed foroptimal synthesis of mRNA. These elements may include splice signals, aswell as transcriptional promoters, enhancers, and termination signals.

Once the expression vector or DNA sequence containing the constructs hasbeen prepared for expression, the expression vectors may be transfectedor introduced into an appropriate host cell. Various techniques may beemployed to achieve this, such as, for example, protoplast fusion,calcium phosphate precipitation, electroporation, retroviraltransduction, viral transfection, gene gun, lipid-based transfection orother conventional techniques. In the case of protoplast fusion, thecells are grown in media and screened for the appropriate activity.Methods and conditions for culturing the resulting transfected cells andfor recovering the antibody molecule produced are known to those skilledin the art and may be varied or optimized depending upon the specificexpression vector and mammalian host cell employed, based upon thepresent description.

In certain embodiments, the host cell comprises a nucleic acid encodingan anti-TIM-3 antibody molecule described herein. In other embodiments,the host cell is genetically engineered to comprise a nucleic acidencoding the anti-TIM-3 antibody molecule.

In one embodiment, the host cell is genetically engineered by using anexpression cassette. The phrase “expression cassette,” refers tonucleotide sequences, which are capable of affecting expression of agene in hosts compatible with such sequences. Such cassettes may includea promoter, an open reading frame with or without introns, and atermination signal. Additional factors necessary or helpful in effectingexpression may also be used, such as, for example, an induciblepromoter. In certain embodiments, the host cell comprises a vectordescribed herein.

The cell can be, but is not limited to, a eukaryotic cell, a bacterialcell, an insect cell, or a human cell. Suitable eukaryotic cellsinclude, but are not limited to, Vero cells, HeLa cells, COS cells, CHOcells, HEK293 cells, BHK cells and MDCKII cells. Suitable insect cellsinclude, but are not limited to, Sf9 cells.

In some embodiments, the host cell is a eukaryotic cell, e.g., amammalian cell, an insect cell, a yeast cell, or a prokaryotic cell,e.g., E. coli. For example, the mammalian cell can be a cultured cell ora cell line. Exemplary mammalian cells include lymphocytic cell lines(e.g., NSO), Chinese hamster ovary cells (CHO), COS cells, oocyte cells,and cells from a transgenic animal, e.g., mammary epithelial cell.

EXAMPLES Example 1

This Example discloses a randomized, double-blind, placebo-controlled,multi-center phase III study design of MBG453 or placebo added toazacitidine for the treatment of subjects with intermediate, high orvery high risk MDS as per IPSS-R or with CMML-2.

Subjects will be randomized in a 1:1 ratio to receive azacitidine 75mg/m2, intravenous or subcutaneous, with or without MBG453 800 mg IV Q4Win 28-day treatment cycles. The randomization will be stratified into 4groups: intermediate risk MDS, high risk MDS, very high risk MDS, andCMML-2. Crossover between treatment arms will not be permitted at anytime during the study.

Study treatment consists of cycles of MBG453 or placebo 800 mg IV Q4Wadministered on Day 8 of each cycle in combination with azacitidineadministered to the subjects on days 1 to 7 (or on days 1 to 5 and days8 and 9) of each cycle until treatment discontinuation. The plannedduration of a cycle is 28 days.

Subjects who become eligible for hematopoietic stem cell transplant(HSCT) or intensive chemotherapy at any time during the course of thestudy may be discontinued from study treatment.

The proposed MBG453 dose in the study is 800 mg Q4W based on dataaccumulated from two phase I studies: [CMBG453X2101] in solid tumorpatients has a wide MBG453 dose range (single agent MBG453 from 80 to1200 mg every 2 weeks (Q2W) or every 4 weeks (Q4W), with a lower 20 mgQ2W MBG453 dose additionally tested in combination with PDR001. Becauseof the data obtained in [CMBG453X2101], study [CPDR001X2105] startedevaluating MBG453 at 240 mg Q2W and additionally tested 400 mg Q2W and800 mg Q4W in combination with decitabine.

The pharmacokinetics (PK) of MBG453 were similar between studies[CMBG453X2101] in solid tumor patients and [CPDR001X2105] in AML andhigh risk MDS patients. At lower doses (at 80 mg and below for Q2Wdosing or at 240 mg and below for Q4W dosing), the PK was nonlinear,with faster elimination at lower concentrations. PK appeared linear withan approximate proportional dose-exposure (AUC and Cmax) relationship atdoses of 240 mg and above for Q2W dosing and at doses of 800 mg andabove for Q4W dosing. Accumulation of MBG453 was observed with repeatedadministrations, and for the Q2W regimen, AUCtau during cycle 3 rangedbetween 1.01-2.78 fold higher than during cycle 1. The dose of 800 mgQ4W has similar AUCtau as 400 mg Q2W at the steady state. In study[CPDR001X2105], clinical benefit was seen across 3 dose levels tested at240 mg Q2W, 400 mg Q2W and 800 mg Q4W in combination with decitabine,with CR or marrow CR in high risk MDS subjects and CR or CRi in newlydiagnosed AML subjects.

Among responding subjects, there were durable responses as long as 19months (as of cut-off date of 25 Mar. 2019). No dose-responserelationship was observed. In a preliminary exposure-response analysis,there was also no clear relationship between exposure and response,using steady state exposure metrics of AUCtau or Ctrough and efficacymetrics of clinical benefit (CR/mCR/CRi) or percent blast cellreduction.

In study [CMBG453X2101], as of 25 Jul. 2019, a total of 133 subjectswith solid tumors have been treated with MBG453 single agent therapy.There were no adverse events attributed to study treatment with anincidence>10%. The most frequently reported adverse events attributed tostudy treatment included fatigue (9%), followed by decreased appetiteand nausea (4.5% each). There were no DLTs during the first cycle. Nosubjects discontinued study treatment due to treatment-related AEs.

In study [CPDR001X2105], as of 26 Jul. 2019, a total of 123 subjectswith hematological malignancies have been treated with MBG453 as asingle agent (n=26) or in combination with decitabine (n=81) orazacitidine (n=16). In the 26 subjects treated with MBG453 single agent,there were no adverse events attributed to study treatment with anincidence>10%. The most frequently reported adverse event attributed tostudy treatment was a rash in two subjects (8%). All other adverseevents attributed to study treatment were single occurrences. There wereno DLTs during the first cycle. No subjects discontinued study treatmentdue to treatment-related AEs. In the 81 subjects treated with MBG453 incombination with decitabine, the most frequent adverse events (allgrades, >10%) attributed to study treatment have includedthrombocytopenia, anemia, neutropenia, nausea, and fatigue. One subjectexperienced a DLT during the first 2 cycles, which consisted ofhepatitis manifesting as Grade 3 ALT increase. One subject discontinuedstudy treatment due to a treatment-related AE of possible hemophagocyticlymphohistiocytosis. In the 16 subjects treated with MBG453 incombination with azacitidine, the most frequent adverse events (allgrades, >10%) attributed to study treatment have included nausea,vomiting, anemia, constipation, neutrophil count decrease, plateletcount decrease. There were no DLTs during the first 2 cycles. Nosubjects discontinued study treatment due to treatment-related AEs. Nostudy treatment-related deaths were observed in any of the studiesmentioned above.

Preliminary analysis revealed no relationship between dose, incidenceand severity of adverse events across the different treatment groups. Norelationship was observed between Cmax and the incidence of potentiallyimmune related adverse events, providing additional support for 800 mgQ4W regimen which has the highest Cmax among the doses tested.

Predicted target engagement: A population pharmacokinetic model ofMBG453 concentration was fit to all subjects from both studies to thepredicted TIM-3 occupancy in the bone marrow by making assumptions aboutMBG453 biodistribution to the bone marrow and binding to TIM-3. Usingtrial simulation, this model predicted that the 800 mg Q4W dose wouldgive at least 95% receptor occupancy in at least 95% of subjects atsteady state Ctrough. This high degree of target engagement is alsosupported by a plateau in the accumulated soluble TIM-3 that is observedat doses of 240 mg Q2W and above, and at 800 mg Q4W and above.

In summary, given the excellent safety and tolerability seen across alldoses and schedules in [CMBG453X2101] and [CPDR001X2105], the activityseen at all 3 doses tested in study [CPDR001X2105]; the predictedsaturation of TIM-3 from the soluble TIM-3 data and the receptoroccupancy model; and the lack of a clear dose-response orexposure-response relationship for MBG453, 800 mg Q4W was selected asthe dose regimen for this study.

Example 2

This example describes the efficacy and safety of sabatolimab (alsoknown as MBG453) in combination with hypomethylating agents (HMAs) inpatients with acute myeloid leukemia (AML) and high-risk myelodysplasticsyndrome (HR-MDS).

Study Design and Methods: This is a phase Ib, open-label, multicenter,dose-escalation study of sabatolimab+HMA (decitabine [Dec] orazacitidine [Aza]) in patients (pts) with AML or HR-MDS (NCT03066648;CPDR001X2105). Patients were adults with newly diagnosed (ND) orrelapsed/refractory (R/R; ≥1 prior therapy) AML or IPSS-R high- or veryhigh-risk MDS; patients with chronic myelomonocytic leukemia (CMML) werealso eligible. Patients were HMA naive and ineligible for intensivechemotherapy. Escalating dose cohorts of IV sabatolimab examined were:240 or 400 mg Q2W (D8, D22) or 800 mg Q4W (D8) combined with Dec (20mg/m²; IV D1-5) or Aza (75 mg/m²; IV/SC D1-7) per 28-day cycle. Primaryobjectives included safety/tolerability; secondary objectives includedpreliminary efficacy and pharmacokinetics.

Results: As of the data cutoff (25 Jun. 2020), 48 patients with ND AML,39 patients with HR-MDS, and 12 patients with CMML receivedsabatolimab+HMA. Data from 29 patients with R/R AML were previouslyreported. For a broader understanding of the effect of sabatolimab+HMA,results are reported here for the Dec and Aza arms both combined andseparately (Table 13). Median (range) duration of sabatolimab exposurewas 4.5 (0.3-28.3) months for ND AML and 4.1 (0.7-33.6) months forHR-MDS, with 17 and 11 patients ongoing, respectively.

With sabatolimab+HMA, the most common (>10% in either disease cohort)grade≥3 treatment-emergent adverse events (TEAEs) in patients with NDAML and HR-MDS, respectively, were thrombocytopenia (45.8%, 51.2%),neutropenia (50%, 46.1%), febrile neutropenia (29.2%, 41%), anemia(27.1%, 28.2%), and pneumonia (10.4%, 5.1%). Discontinuation due to AEwas infrequent among patients with ND AML (6.3% [3/48]; 1 each offatigue, febrile neutropenia, and possible HLH); none occurred amongpatients with HR-MDS. One dose-limiting toxicity occurred withsabatolimab 240 mg Q2W+Dec (grade 3 ALT elevation); the maximumtolerated dose was not reached with either combination.

To comprehensively assess possible immune-mediated AEs (imAEs), eventswere evaluated across all disease cohorts. Seven grade 3treatment-related possible imAEs were reported in 5 patients (increasedALT [2 patients], and arthritis, possible HLH, infusion-relatedreaction, hypothyroidism, and rash [1 patient each]). No grade 4treatment-related possible imAEs occurred; however, there was a case ofenterocolitis in a patient with HR-MDS who died of septic shock withneutropenic colitis. No other treatment-related deaths were reported.

Among 34 evaluable patients with ND AML, overall response rate (ORR) was41.2%: 8 CR, 3 CRi, 3 PR. Median (range) time to response (TTR) was 2.1(1.8-13.1) months and estimated 6-month duration of response (DOR) ratewas 85.1% (95% CI: 68-100%). Estimated 12-month progression-freesurvival (PFS) rate was 44% (95% CI: 28-69.3%). Among 35 evaluablepatients with HR-MDS, ORR was 62.9%: 8 CR, 8 mCR, (5 with hematologicimprovement MID, 6 SD+HI. Median (range) TTR was 2.0 (1.7-9.6) monthsand estimated 6-month DOR rate for CR/mCR/PR was 90% (95% CI:73.2-100%). Encouraging response rates were achieved in both patientswith high-risk MDS (ORR 50% [11/22]) and very high-risk MDS (ORR 84.6%[11/13]). Of patients with HR-MDS, 8 (5 high-risk, 3 very high-risk)proceeded to transplant. Estimated 12-month PFS rate was 58.1% (95% CI:39.9-84.6%).

Among 12 patients with CMML, the safety profile of sabatolimab+HMA wasgenerally consistent with that for AML/HR-MDS (most common grade ≥3TEAEs: thrombocytopenia, n=7; neutropenia, n=7; anemia, n=6). ORR among11 evaluable patients was 63.6%: 2 CR, 3 mCR, 1 PR, 1 SD+HI.

Conclusions: Sabatolimab+HMA is well tolerated in patients with AML andHR-MDS and continues to show promising antileukemic activity andemerging durability. These results support TIM-3 as a potentialtherapeutic target and provide a basis for further development ofsabatolimab+HMA in patients with AML or higher-risk MDS.

TABLE 13 Summary of results of following administration of sabatolimab +HMA to patients with newly diagnosed (ND) AML, high-risk (HR) MDS, orCMML ND AML HR-MDS CMML^(a) + Dec + Aza + Dec + Aza + Dec + AzaParameter n = 22 n = 26 n = 19 n = 20 n = 5 n = 7 Duration ofsabatolimab 6.8 3.5 8.0 2.8 8.4 5.0 exposure, median (range) (0.7-28.3)(0.3-15.2) (0.7-33.6) (0.8-14.3) (5.6-12.6) (1.6-15.8) months Efficacyevaluable patients, 17 17 18 17 5 6 n ORR^(b), n (%) 8 (47.1) 6 (35.3)11 (61.1) 11 (64.7) 3 (60) 4 (66.7) CR 6 (35.3 2 (11.8) 6 (33.3) 2(11.8) 0 2 (33.3) CRi 1 (5.9) 2 (11.8) NA NA NA NA mCR NA NA 3 (16.7) 5(29.4) 1 (20) 2 (33.3) mCR with HI NA NA 3 (16.7) 2 (11.8) 0 1 (16.7) PR1 (5.9) 2 (11.8) 0 0 1 (20) 0 SD with HI NA NA 2 (11.1) 4 (23.5) 1 (20)0 ^(a)Response assessment for patients with CMML used IWG criteria(Cheson 2006). ^(b)ORR for patients with MDS was defined as CR + mCR +PR + SD with HI; ORR for patients with ND AML was defined as CR + CRi +PR. CR, complete remission; CRi, CR with incomplete blood countrecovery; mCR, marrow CR; PR, partial remission.

Example 3—MBG453 Partially Blocks the Interaction Between TIM-3 andGalectin 9

Galectin-9 is a ligand of TIM-3. Asayama et al. (Oncotarget 8(51):88904-88971 (2017) demonstrated by the TIM-3-Galectin 9 pathway isassociated with the pathogenesis and disease progression of MDS. Thisexample illustrates the ability of MBG453 to partially block theinteraction between TIM-3 and Galectin 9.

TIM-3 fusion protein (R&D Systems) was coated on a standard MesoScale 96well plate (Meso Scale Discovery) at 2 μg/ml in PBS (Phosphate BufferedSaline) and incubated for six hours at room temperature. The plate waswashed three times with PBST (PBS buffer containing 0.05% Tween-20) andblocked with PBS containing 5% Probumin (Millipore) overnight at 4° C.After incubation, the plate was washed three times with PBST andunlabeled antibody (F38-2E2 (BioLegend); MBG453; MBG453 F(ab′)2; MBG453F(ab); or control recombinant human Galectin-9 protein) diluted in AssayDiluent (2% Probumin, 0.1% Tween-20, 0.1% Triton X-100 (Sigma) with 10%StabilGuard (SurModics)), was added in serial dilutions to the plate andincubated for one hour on an orbital shaker at room temperature. Theplate was then washed three times with PBST, and Galectin-9 labeled withMSD SULFOTag (Meso Scale Discovery) as per manufacturer's instructions,diluted in Assay Diluent to 100 nM, was added to the plate for one hourat room temperature on an orbital shaker. The plate was again washedthree times with PBST, and Read Buffer T (1×) was added to the plate.The plate was read on MA600 Imager, and competition was assessed as ameasure of the ability of the antibody to block Gal9-SULFOTag signal toTIM-3 receptor. As shown in FIG. 1 , MBG453 IgG4, MBG453 F(ab′)2, MBG453F(ab), and 2E2 partially blocked the interaction between TIM-3 andGalectin-9, whereas control Galectin-9 protein did not.

Example 4—MBG453 Mediates Antibody-Dependent Cellular Phagocytosis(ADCP) Through Engagement of FcγR1

THP-1 effector cells (a human monocytic AML cell line) weredifferentiated into phagocytes by stimulation with 20 ng/ml phorbol12-myristate 13-acetate (PMA) for two to three days at 37° C., 5% CO2.PMA-stimulated THP-1 cells were washed in FACS Buffer (PBS with 2 mMEDTA) in the flask and then detached by treatment with Accutase(Innovative Cell Technologies). The target TIM-3-overexpressing Rajicells were labelled with 5.5 μM CellTrace CFSE (ThermoFisherScientific)as per manufacturer's instructions. THP-1 cells and TIM-3-overexpressingCFSE+ Raji cells were co-cultured at an effector to target (E:T) ratioof 1:5 with dilutions of MBG453, MabThera anti-CD20 (Roche) positivecontrol, or negative control antibody (hIgG4 antibody with target notexpressed by the Raji TIM-3+ cells) in a 96 well plate (spun at 100×gfor 1 minute at room temperature at assay start). Co-cultures wereincubated for 30-45 minutes at 37° C., 5% CO2. Phagocytosis was thenstopped with a 4% Formaldehyde fixation (diluted from 16% stock,ThermoFisherScientific), and cells were stained with an APC-conjugatedanti-CD11c antibody (BD Bioscience). ADCP was measured by a flowcytometry based assay on a BD FACS Canto II. Phagocytosis was evaluatedas a percentage of the THP-1 cells double positive for CFSE(representing the phagocytosed Raji cell targets) and CD11c from theTHP-1 (effector) population. As shown in FIG. 2 , MBG453 (squares)enhanced THP-1 cell phagocytosis of TIM-3+ Raji cells in adose-dependent manner, which then plateaued relative to the anti-CD20positive control (open circles). Negative control IgG4 antibody is shownin triangles.

The TIM-3-expressing Raji cells were used as target cells in aco-culture assay with engineered effector Jurkat cells stablytransfected to overexpress FcγR1a (CD64) and a luciferase reporter geneunder the control of an NFAT (nuclear factor of activated T cells)response element (NFAT-RE; Promega). The target TIM-3+ Raji cells wereco-incubated with the Jurkat-FcγR1a reporter cells in an E:T ratio of6:1 and graded concentrations (500 ng/ml to 6 pg/ml) of MBG453 or theanti-CD20 MabThera reference control (Roche) in a 96 well plate. Theplate was then centrifuged at 300×g for 5 minutes at room temperature atthe assay start and incubated for 6 hours in a 37° C., 5% CO₂ humidifiedincubator. The activation of the NFAT dependent reporter gene expressioninduced by the binding to FcγR1a was quantified by luciferase activityafter cell lysis and the addition of a substrate solution (Bio-GLO). Asshown in FIG. 3 , MBG453 showed a modest dose-response engagement of theFcγR1a reporter cell line as measured by luciferase activity. In aseparate assay, MBG453 did not engage FcγRIIa (CD32a).

Example 5—MBG453 Enhances Immune-Mediated Killing of DecitabinePre-Treated AML Cells

THP-1 cells were plated in complete RPMI-1640 (Gibco) media(supplemented with 2 mM glutamine, 100 U/ml Pen-Strep, 10 mM HEPES, 1 mMNaPyr, and 10% fetal bovine serum (FBS)). Decitabine (250 or 500 nM;supplemented to media daily for five days) or DMSO control were addedfor a 5-day incubation at 37° C., 5% CO₂. Two days after plating THP-1cells, healthy human donor peripheral blood mononuclear cells (PBMCs;Medcor) were isolated from whole blood by centrifugation of sodiumcitrate CPT tubes at 1,800×g for 20 minutes. At the completion of thespin, the tube was inverted 10 times to mix the plasma and PBMC layers.Cells were washed in 2× volume of PBS/MACS Buffer (Miltenyi) andcentrifuged at 250×g for 5 minutes. Supernatant was aspirated, and 1 mLof PBS/MACS Buffer was added following by pipetting to wash the cellpellet. 19 mL of PBS/MACS Buffer were added to wash, followed by arepeat of the centrifugation. Supernatant was aspirated, and the cellpellet was resuspended in 1 mL of complete media, followed by pipettingto a single cell suspension, and the volume was brought up to 10 mL withcomplete RPMI. 100 ng/mL anti-CD3 (eBioscience) was added to the mediafor a 48-hour stimulation at 37° C., 5% CO₂. After 5 days culture withdecitabine or DMSO, THP-1 cells were harvested and labeled withCellTracker™ Deep Red Dye (ThermoFisher) following manufacturer'sinstructions.

Labeled THP-1 cells (decitabine pre-treated or DMSO control-treated)were co-cultured with stimulated PBMCs at effector:target (E:T) ratiosof 1:1, 1:2, and 1:3 (optimized for each donor, with the target cellnumber constant at 10,000 cells/well (Costar 96 well flat bottom plate).Wells were treated with either hIgG4 isotype control or MBG453 at 1μg/mL. The plate was placed in an Incucyte S3, and image phase and redfluorescent channels were captured every 4 hours for 5 days. At thecompletion of the assay, the target cell number (red events) wasnormalized to the first imaging time point using the Incucyte imageanalysis software.

As shown in FIG. 4 , co-culture of THP-1 cells with anti-CD3 activatedPBMCs led to killing of the THP-1 cells, enhanced in the presence ofMBG453 (bars in bottom violin plot, each dot represents a single healthyPBMC donor) relative to hIgG4 isotype control at the terminal timepointof the assay. This killing was further enhanced by pre-treatment of theTHP-1 cells with decitabine (bars in top violin plot, each dotrepresents a single healthy PBMC donor). Taken together, these dataindicate that MBG453 blockade of TIM-3 enhanced immune-mediated killingof THP-1 AML cells, with pre-treatment with decitabine further enhancingthis activity.

Example 6—Investigation of MBG453 and Decitabine-Mediated Killing ofPatient-Derived Xenografts in an Immuno-Deficient Host

The activity of MBG453 with and without decitabine was evaluated in twoAML patient-derived xenograft (PDX) models, HAMLX21432 and HAMLX5343.Decitabine (TCI America) was formulated in dextrose 5% in water (D5W)freshly prior to each dose and administered daily for 5 days. It wasadministered at 10 mL/kg intraperitoneal (i.p.), for a final dose volumeof 1 mg/kg. MBG453 was formulated to a final concentration of 1 mg/mL inPBS. It was administered weekly at a volume of 10 mL/kg, i.p., for afinal dose of 10 mg/kg, with treatment initiating on dosing day 6, 24hours after the final dose of decitabine. The combination of MBG453 anddecitabine was well-tolerated as measured both by body weight changemonitoring and visual inspection of health status in both models.

For one study, mice were injected with 2×10⁶ cells intravenously (i.v.)that were isolated from an in vivo passage 5 of the AML PDX #21432 modelharboring an IDH1R132H mutation. Animals were randomized into treatmentgroups once they reached a leukemic burden on average of 39%. Treatmentswere initiated on the day of randomization and continued for 21 daysAnimals remained on study until each reached individual endpoints,defined by circulating leukemic burden of greater than 90% human CD45+cells, body weight loss>20%, signs of hind limb paralysis, or poor bodycondition. HAML21432 implanted mice treated with decitabine alonedemonstrated moderate anti-tumor activity that peaked at approximatelyday 49 post-implant or day 14 post-treatment start. At this time point,decitabine-treated groups were on average at 51% and 47% hCD45+ cells,single agent and combination with MBG453, respectively (FIG. 5 ). At thesame time point, the untreated and MBG453-treated groups were at aleukemic burden of 81% and 77%, respectively. By day 56post-implantation, however, the decitabine-treated groups increased inleukemic burden to 66% and 61% hCD45+ cells in circulation. Nocombination activity was observed when decitabine was combined withMBG453 in this model (FIG. 5 ). Untreated and MBG453 single agenttreated groups both reached the time to end point cut off of 90%leukemic burden by day 56.

For another study, mice were injected with 2×10⁶ cells i.v. that wereisolated from an in vivo passage 4 of the AML PDX #5343 model harboringmutations KRASG12D, WT1 and PTPN11. Animals were randomized intotreatment groups once they reached a leukemic burden on average of 20%.Treatments were initiated on the day of randomization and continued for3 weeks Animals remained on study until each reached individualendpoints, defined by circulating leukemic burden of greater than 90%human CD45+ cells, body weight loss>20%, signs of hind limb paralysis orpoor body condition. HAML5343 implanted mice treated with decitabinealone showed significant anti-tumor activity with a peak ofapproximately day 53 post-implant or day 21 post-treatment start. Atthis time point, decitabine-treated groups were on average at 1% and1.3% hCD45+ cells, single agent and combination with MBG453,respectively (FIG. 6 ). At the same time point, the untreated group hada leukemic burden of 91%. The MBG453-treated group only had oneremaining animal by day 53. No combination activity was observed whendecitabine was combined with MBG453 in this model (FIG. 6 ). Thesignificant reduction in tumor burden was comparable in decitabinesingle agent and decitabine/MBG453 combination groups in this model.

The Nod scid gamma (NSG; NOD.Cg-prkdc<scid>Il2rg<tm1wj1>/SzJ, Jackson)model used for the AML PDX implantation lacks immune cells, likely suchas TIM-3-expressing T cells, NK cells, and myeloid cells, indicatingcertain immune cell functions may be required for MBG453 to enhance theactivity of decitabine in the mouse model.

Example 7—MBG453 Enhances Killing of Thp-1 AML Cells that are Engineeredto Overexpress TIM-3

THP-1 cells express TIM-3 mRNA but low to no TIM-3 protein on the cellsurface. THP-1 cells were engineered to stably overexpress TIM-3 with aFlag-tag encoded by a lentiviral vector, whereas parental THP-1 cells donot express TIM-3 protein on the surface. TIM-3 Flag-tagged THP-1 cellswere labeled with 2 μM CFSE (Thermo Fisher Scientific), and THP-1parental cells were labeled with 2 μM CTV (Thermo Fisher Scientific),according to manufacturer instructions. Co-culture assays were performedin 96-well round-bottom plates. THP-1 cells were mixed at a 1:1 ratiofor a total of 100,000 THP-1 cells per well (50,000 THP-1 expressingTIM-3 and 50,000 THP-1 parental cells) and co-cultured for three dayswith 100,000 T cells purified using a human pan T cell isolation kit(Miltenyi Biotec) from healthy human donor PBMCs (Bioreclamation), inthe presence of varying amounts of anti-CD3/anti-CD28 T cell activationbeads (ThermoFisherScientific) and 25 μg/ml MBG453 (whole antibody),MBG453 F(ab), or hIgG4 isotype control. Cells were then detected andcounted by flow cytometry. The ratio between TIM-3-expressing THP-1cells and parental THP-1 cells (“fold” in y-axis of graph) wascalculated and normalized to conditions without anti-CD3/anti-CD28 beadstimulation. The x-axis of the graph denotes the stimulation amount asnumber of beads per cell. Data represents one of two independentexperiments. As seen in FIG. 7 , MBG453 (triangles) but not MBG453 F(ab)(open squares) enhances the T cell-mediated killing of THP-1 cells thatoverexpress TIM-3 relative to parental control THP-1 cells indicatingthat the Fc-portion of MBG453 can be important for MBG453-enhanced Tcell-mediated killing of THP-1 AML cells.

Embodiments of the Application

The following are embodiments disclosed in the present application. Theembodiments include, but are not limited to:

1. A combination comprising a TIM-3 inhibitor and a hypomethylatingagent for use in treating a myelodysplastic syndrome (MDS) or a chronicmyelomonocytic leukemia (CMML), in a subject.

2. A method of treating a myelodysplastic syndrome (MDS) or a chronicmyelomonocytic leukemia (CMML), in a subject, comprising administeringto the subject a combination of a TIM-3 inhibitor and hypomethylatingagent. 3. The combination for use of embodiment 1, or the method ofembodiment 2, wherein the TIM-3 inhibitor comprises an anti-TIM-3antibody molecule.

4. The combination for use of embodiment 1 or 3, or the method ofembodiment 2 or 3, wherein the TIM-3 inhibitor comprises MBG453 orTSR-022.

5. The combination for use of embodiment 1 or 3, or the method ofembodiment 2 or 3, wherein the TIM-3 inhibitor comprises MBG453.

6. The combination for use of any of embodiments 1 or 3-5, or the methodof any of embodiments 2-5, wherein the TIM-3 inhibitor is administeredat a dose of about 700 mg to about 900 mg.

7. The combination for use of any of embodiments 1 or 3-6, or the methodof any of embodiments 2-6, wherein the TIM-3 inhibitor is administeredat a dose of about 800 mg.

8. The combination for use of any of embodiments 1 or 3-7, or the methodof any of embodiments 2-7, wherein the TIM-3 is administered at day 8 ofa 28-day cycle.

9. The combination for use of any of embodiments 1 or 3-8, or the methodof any of embodiments 2-8, wherein the TIM-3 inhibitor is administeredonce every four weeks. 10. The combination for use of any of embodiments1 or 3-9, or the method of any of embodiments 2-9, wherein the TIM-3inhibitor is administered intravenously.

11. The combination for use of any of embodiments 1 or 3-10, or themethod of any of embodiments 2-10, wherein the TIM-3 inhibitor isadministered intravenously over a period of about 15 minutes to about 45minutes.

12. The combination for use of any of embodiments 1 or 3-11, or themethod of any of embodiments 2-11, wherein the TIM-3 inhibitor isadministered intravenously over a period of about 30 minutes.

13. The combination for use of embodiments 1 or 3-12, or the method ofembodiments 2-12, wherein the hypomethylating agent comprisesazacitidine or decitabine.

14. The combination for use of embodiments 1 or 3-13, or the method ofembodiments 2-13, wherein the hypomethylating agent comprisesazacitidine.

15. The combination for use of any of embodiments 1 or 3-14, or themethod of any of embodiments 2-14, wherein the hypomethylating agent isadministered at a dose of about 50 mg/m² to about 100 mg/m².

16. The combination for use of any of embodiments 1 or 3-15, or themethod of any of embodiments 2-15, wherein the hypomethylating agent isadministered at a dose of about 75 mg/m².

17. The combination for use of any of embodiments 1 or 3-16, or themethod of any of embodiments 2-16, wherein the hypomethylating agent isadministered once a day.

18. The combination for use of any of embodiments 1 or 3-17, or themethod of any of embodiments 2-17, wherein the hypomethylating agent isadministered for 5-7 consecutive days.

19. The combination for use of any of embodiments 1 or 3-18, or themethod of any of embodiments 2-18, wherein the hypomethylating agent isadministered for (a) seven consecutive days on days 1-7 of a 28-daycycle, or (b) five consecutive days on days 1-5, followed by a two-daybreak, then two consecutive days on days 8-9, of a 28-day cycle.

20. The combination for use of any of embodiments 1 or 3-19, or themethod of any of embodiments 2-19, wherein the hypomethylating agent isadministered subcutaneously or intravenously.

21. The combination for use of any of embodiments 1 or 3-20, or themethod of any of embodiments 2-20, wherein the myelodysplastic syndrome(MDS) is an intermediate MDS, high risk MDS, or very high risk MDS.

22. The combination for use of any of embodiments 1 or 3-20, or themethod of any of embodiments 2-20, wherein the chronic myelomonocyticleukemia (CMML) is CMML-1 or CMML-2.

23. A combination comprising MBG453 and azacitidine for use in treatinga CMML-2 in a subject.

24. A combination comprising MBG453 and azacitidine for use in treatingan intermediate MDS, high risk MDS, or very high risk MDS in a subject.

25. A method of treating a CMML-2 in a subject, comprising administeringto the subject a combination of MBG453 and azacitidine.

26. A method of treating an intermediate MDS, high risk MDS, or veryhigh risk MDS in a subject, comprising administering to the subject acombination of MBG453 and azacitidine.

27. The combination for use of embodiment 23 or 24, or the method ofembodiment 25 or 26, wherein MBG453 is administered at a dose of about700 mg to about 900 mg.

28. The combination for use of embodiment 23-24 or 27, or the method ofembodiment 25-27, wherein MBG453 is administered at a dose of about 800mg.

29. The combination for use of any of embodiments 23-24 or 27-28, or themethod of any of embodiments 25-28, wherein MBG453 is administered onceevery four weeks.

30. The combination for use of any of embodiments 23-24 or 27-29, or themethod of any of embodiments 25-29, wherein MBG453 is administered atday 8 of a 28-day cycle.

31. The combination for use of any of embodiments 23-24 or 27-30, or themethod of any of embodiments 25-30, wherein MBG453 is administered onceevery four weeks.

32. The combination for use of any of embodiments 23-24 or 27-31, or themethod of any of embodiments 25-31, wherein MBG453 is administeredintravenously

33. The combination for use of any of embodiments 23-24 or 27-32, or themethod of any of embodiments 25-32, wherein MBG453 is administeredintravenously over a period of about 15 minutes to about 45 minutes.

34. The combination for use of any of embodiments 23-24 or 27-33, or themethod of any of embodiments 25-33, wherein MBG453 is administeredintravenously over a period of about 30 minutes.

35. The combination for use of any of embodiments 23-24 or 27-34, or themethod of any of embodiments 25-34, wherein azacitidine is administeredat a dose of about 50 mg/m² to about 100 mg/m².

36. The combination for use of any of embodiments 23-24 or 27-35, or themethod of any of embodiments 25-35, wherein azacitidine is administeredat a dose of about 75 mg/m².

37. The combination for use of any of embodiments 23-24 or 27-36, or themethod of any of embodiments 25-36, wherein azacitidine is administeredonce a day.

38. The combination for use of any of embodiments 23-24 or 27-37, or themethod of any of embodiments 25-37, wherein azacitidine is administeredfor 5-7 consecutive days.

39. The combination for use of any of embodiments 23-24 or 27-38, or themethod of any of embodiments 25-38, wherein azacitidine is administeredfor (a) seven consecutive days on days 1-7 of a 28-day cycle, or (b)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.

40. The combination for use of any of embodiments 23-24 or 27-39, or themethod of any of embodiments 25-39, wherein azacitidine is administeredsubcutaneously or intravenously.

41. A method of treating a CMML-2 in a subject, comprising administeringto the subject a combination of MBG453 and azacitidine, wherein:

a) MBG453 is administered at a dose of about 800 mg once every fourweeks on day 8 of a 28-day dosing cycle; and

b) azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.

42. A combination comprising MBG453 and azacitidine for use in treatinga CMML-2 in a subject, wherein:

a) MBG453 is administered at a dose of about 800 mg once every fourweeks on day 8 of a 28-day dosing cycle; and

b) azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.

43. A method of treating an intermediate MDS, a high risk MDS, or a veryhigh risk MDS in a subject, comprising administering to the subject acombination of MBG453 and azacitidine, wherein:

a) MBG453 is administered at a dose of about 800 mg once every fourweeks on day 8 of a 28-day dosing cycle; and

b) azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.

44. A combination comprising MBG453 and azacitidine for use in treatingan intermediate MDS, a high risk MDS, or a very high risk MDS in asubject, wherein:

a) MBG453 is administered at a dose of about 800 mg once every fourweeks on day 8 of a 28-day dosing cycle; and

b) azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.

INCORPORATION BY REFERENCE

All publications, patents, and Accession numbers mentioned herein arehereby incorporated by reference in their entirety as if each individualpublication or patent was specifically and individually indicated to beincorporated by reference.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification and the claims below. The fullscope of the invention should be determined by reference to the claims,along with their full scope of equivalents, and the specification, alongwith such variations.

What is claimed is:
 1. A combination comprising a TIM-3 inhibitor and ahypomethylating agent for use in treating a myelodysplastic syndrome(MDS) or a chronic myelomonocytic leukemia (CMML), in a subject.
 2. Amethod of treating a myelodysplastic syndrome (MDS) or a chronicmyelomonocytic leukemia (CMML), in a subject, comprising administeringto the subject a combination of a TIM-3 inhibitor and hypomethylatingagent.
 3. The combination for use of claim 1, or the method of claim 2,wherein the TIM-3 inhibitor comprises an anti-TIM-3 antibody molecule.4. The combination for use of claim 1 or 3, or the method of claim 2 or3, wherein the TIM-3 inhibitor comprises MBG453 or TSR-022.
 5. Thecombination for use of claim 1 or 3, or the method of claim 2 or 3,wherein the TIM-3 inhibitor comprises MBG453.
 6. The combination for useof any of claim 1 or 3-5, or the method of any of claims 2-5, whereinthe TIM-3 inhibitor is administered at a dose of about 700 mg to about900 mg.
 7. The combination for use of any of claim 1 or 3-6, or themethod of any of claims 2-6, wherein the TIM-3 inhibitor is administeredat a dose of about 800 mg.
 8. The combination for use of any of claim 1or 3-7, or the method of any of claims 2-7, wherein the TIM-3 isadministered at day 8 of a 28-day cycle.
 9. The combination for use ofany of claim 1 or 3-8, or the method of any of claims 2-8, wherein theTIM-3 inhibitor is administered once every four weeks.
 10. Thecombination for use of any of claim 1 or 3-9, or the method of any ofclaims 2-9, wherein the TIM-3 inhibitor is administered intravenously.11. The combination for use of any of claim 1 or 3-10, or the method ofany of claims 2-10, wherein the TIM-3 inhibitor is administeredintravenously over a period of about 15 minutes to about 45 minutes. 12.The combination for use of any of claim 1 or 3-11, or the method of anyof claims 2-11, wherein the TIM-3 inhibitor is administeredintravenously over a period of about 30 minutes.
 13. The combination foruse of claim 1 or 3-12, or the method of claims 2-12, wherein thehypomethylating agent comprises azacitidine or decitabine.
 14. Thecombination for use of claim 1 or 3-13, or the method of claims 2-13,wherein the hypomethylating agent comprises azacitidine.
 15. Thecombination for use of any of claim 1 or 3-14, or the method of any ofclaims 2-14, wherein the hypomethylating agent is administered at a doseof about 50 mg/m² to about 100 mg/m².
 16. The combination for use of anyof claim 1 or 3-15, or the method of any of claims 2-15, wherein thehypomethylating agent is administered at a dose of about 75 mg/m². 17.The combination for use of any of claim 1 or 3-16, or the method of anyof claims 2-16, wherein the hypomethylating agent is administered once aday.
 18. The combination for use of any of claim 1 or 3-17, or themethod of any of claims 2-17, wherein the hypomethylating agent isadministered for 5-7 consecutive days.
 19. The combination for use ofany of claim 1 or 3-18, or the method of any of claims 2-18, wherein thehypomethylating agent is administered for (a) seven consecutive days ondays 1-7 of a 28-day cycle, or (b) five consecutive days on days 1-5,followed by a two-day break, then two consecutive days on days 8-9, of a28-day cycle.
 20. The combination for use of any of claim 1 or 3-19, orthe method of any of claims 2-19, wherein the hypomethylating agent isadministered subcutaneously or intravenously.
 21. The combination foruse of any of claim 1 or 3-20, or the method of any of claims 2-20,wherein the myelodysplastic syndrome (MDS) is an intermediate MDS, highrisk MDS, or very high risk MDS.
 22. The combination for use of any ofclaim 1 or 3-20, or the method of any of claims 2-20, wherein thechronic myelomonocytic leukemia (CMML) is CMML-1 or CMML-2.
 23. Acombination comprising MBG453 and azacitidine for use in treating aCMML-2 in a subject.
 24. A combination comprising MBG453 and azacitidinefor use in treating an intermediate MDS, high risk MDS, or very highrisk MDS in a subject.
 25. A method of treating a CMML-2 in a subject,comprising administering to the subject a combination of MBG453 andazacitidine.
 26. A method of treating an intermediate MDS, high riskMDS, or very high risk MDS in a subject, comprising administering to thesubject a combination of MBG453 and azacitidine.
 27. The combination foruse of claim 23 or 24, or the method of claim 25 or 26, wherein MBG453is administered at a dose of about 700 mg to about 900 mg.
 28. Thecombination for use of claim 23-24 or 27, or the method of claim 25-27,wherein MBG453 is administered at a dose of about 800 mg.
 29. Thecombination for use of any of claim 23-24 or 27-28, or the method of anyof claims 25-28, wherein MBG453 is administered once every four weeks.30. The combination for use of any of claim 23-24 or 27-29, or themethod of any of claims 25-29, wherein MBG453 is administered at day 8of a 28-day cycle.
 31. The combination for use of any of claim 23-24 or27-30, or the method of any of claims 25-30, wherein MBG453 isadministered once every four weeks.
 32. The combination for use of anyof claim 23-24 or 27-31, or the method of any of claims 25-31, whereinMBG453 is administered intravenously.
 33. The combination for use of anyof claim 23-24 or 27-32, or the method of any of claims 25-32, whereinMBG453 is administered intravenously over a period of about 15 minutesto about 45 minutes.
 34. The combination for use of any of claim 23-24or 27-33, or the method of any of claims 25-33, wherein MBG453 isadministered intravenously over a period of about 30 minutes.
 35. Thecombination for use of any of claim 23-24 or 27-34, or the method of anyof claims 25-34, wherein azacitidine is administered at a dose of about50 mg/m² to about 100 mg/m².
 36. The combination for use of any of claim23-24 or 27-35, or the method of any of claims 25-35, whereinazacitidine is administered at a dose of about 75 mg/m².
 37. Thecombination for use of any of claim 23-24 or 27-36, or the method of anyof claims 25-36, wherein azacitidine is administered once a day.
 38. Thecombination for use of any of claim 23-24 or 27-37, or the method of anyof claims 25-37, wherein azacitidine is administered for 5-7 consecutivedays.
 39. The combination for use of any of claim 23-24 or 27-38, or themethod of any of claims 25-38, wherein azacitidine is administered for(a) seven consecutive days on days 1-7 of a 28-day cycle, or (b) fiveconsecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.
 40. The combination foruse of any of claim 23-24 or 27-39, or the method of any of claims25-39, wherein azacitidine is administered subcutaneously orintravenously.
 41. A method of treating a CMML-2 in a subject,comprising administering to the subject a combination of MBG453 andazacitidine, wherein: a) MBG453 is administered at a dose of about 800mg once every four weeks on day 8 of a 28-day dosing cycle; and b)azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.
 42. A combinationcomprising MBG453 and azacitidine for use in treating a CMML-2 in asubject, wherein: a) MBG453 is administered at a dose of about 800 mgonce every four weeks on day 8 of a 28-day dosing cycle; and b)azacitidine is administered at a dose of about 75 mg/m² a day for (i)seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.
 43. A method oftreating an intermediate MDS, a high risk MDS, or a very high risk MDSin a subject, comprising administering to the subject a combination ofMBG453 and azacitidine, wherein: a) MBG453 is administered at a dose ofabout 800 mg once every four weeks on day 8 of a 28-day dosing cycle;and b) azacitidine is administered at a dose of about 75 mg/m² a day for(i) seven consecutive days on days 1-7 of a 28-day dosing cycle, or (ii)five consecutive days on days 1-5, followed by a two-day break, then twoconsecutive days on days 8-9, of a 28-day cycle.
 44. A combinationcomprising MBG453 and azacitidine for use in treating an intermediateMDS, a high risk MDS, or a very high risk MDS in a subject, wherein: a)MBG453 is administered at a dose of about 800 mg once every four weekson day 8 of a 28-day dosing cycle; and b) azacitidine is administered ata dose of about 75 mg/m² a day for (i) seven consecutive days on days1-7 of a 28-day dosing cycle, or (ii) five consecutive days on days 1-5,followed by a two-day break, then two consecutive days on days 8-9, of a28-day cycle.